Uridine nucleoside derivatives, compositions and methods of use

ABSTRACT

This disclosure relates to uridine nucleoside derivatives, compositions comprising therapeutically effective amounts of those nucleoside derivatives and methods of using those nucleoside derivatives or compositions in treating disorders that are responsive to ligands, such as agonists, of P 2 Y 6  receptor, e.g., neuronal disorders, including neurodegenerative disorders (e.g., Alzheimer&#39;s disease, Parkinson&#39;s disease) and traumatic CNS injury, pain, Down Syndrome (DS), glaucoma and inflammatory conditions.

RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No.14/775,426, filed Sep. 11, 2015 (U.S. Pat. No. 10,544,183), which is anational stage filing under 35 U.S.C 371 of International ApplicationNo.: PCT/US2014/026865, filed Mar. 13, 2014, which claims the benefit ofand priority from U.S. Provisional Patent Applications 61/780,219, filedMar. 13, 2013, and 61/883,604, filed Sep. 27, 2013. Each of theforegoing applications is incorporated herein by reference in itsentirety. International Application No.: PCT/US2014/026865 was publishedunder PCT Article 21(2) in English.

FIELD OF THE INVENTION

This disclosure relates to compounds, compositions and methods fortreating neuronal disorders, including neurodegenerative disorders(e.g., Alzheimer's disease, Parkinson's disease) and traumatic CNSinjury, pain, Down Syndrome (DS), glaucoma and inflammatory conditions.

BACKGROUND OF THE INVENTION

P₂Y receptors are G-protein-coupled receptors (GPCRs) that areselectively activated by naturally occurring extracellular nucleotides,including, for example, adenine and pyrimidine nucleotides. There aretwo clusters of P₂Y receptors: the G_(q)-coupled P₂Y₁-like receptors,including P₂Y_(1,2,4,6,11) subtypes; and the G_(i)-coupled P₂Y₁₂-likereceptors, including P₂Y_(12, 13, 14) subtypes. Of the four P₂Yreceptors, i.e., P₂Y_(2, 4, 6, 14) subtypes, which can be activated bypyrimidine nucleotides, the P₂Y₂ and P₂Y₄ subtypes are activated byuridine triphosphate (UTP), P₂Y₆ receptor is activated by uridinediphosphate (UDP), and P₂Y₁₄ is activated by UDP or UDP-glucose.

The P₂Y₆ receptor has been implicated in a number of disorders,including, for example, neurodegeneration, osteoporosis, ischemic effectin skeletal muscle, and diabetes. It has been reported that agonists ofP₂Y₆ receptor counteract apoptosis induced by tumor necrosis factor α inastrocytoma cells and induce protection in a model of ischemic hindlegskeletal muscle. P₂Y₆ receptor was also reported to play a role inphagocytosis in microglial cells when activated by its endogenousagonist UDP. See, e.g., Malmsjo et al. BMC Pharmacol. 2003, 3, 4;Balasubramanian et al. Biochem. Pharmacol. 2010, 79, 1317-1332; Kim etal. Cell. Mol. Neurobiol. 2003, 23, 401-418; Mamedova et al. Pharmacol.Res. 2008, 58, 232-239; Korcok et al. J. Biol. Chem. 2005, 58, 232-239;and Koizumi et al. Nature, 2007, 446, 1091-1095. These reports suggestthat ligands of the P₂Y₆ receptor are of interest in the search for newtreatments for P₂Y₆ receptor-related conditions.

Therefore, there is a need for new ligands, such as agonists, of P₂Y₆receptor activity that are useful in therapeutic preparations for thetreatment of disorders mediated by the receptor, includingneurodegeneration, traumatic brain injury and pain.

SUMMARY OF THE INVENTION

The present disclosure addresses the aforementioned need by providingcompounds of formulae I and II:

wherein the variables are as defined herein, along with pharmaceuticallyacceptable salts thereof. In certain embodiments, these compounds arecapable of modulating P₂Y₆ receptor activity, either directly orindirectly, i.e., these compounds are P₂Y₆ receptor-modulatingcompounds. In certain embodiments, the compounds as described herein areagonists of the P₂Y₆ receptor, which, directly or indirectly, activatethe P₂Y₆ receptor. Compounds of formulae I (including formula I-A andformula I-B) and II can be used to treat the conditions as describedherein.

The present disclosure also provides compositions that comprise thecompounds described herein. The disclosure also includes the use of thecompounds disclosed herein in the manufacture of a medicament for thetreatment of one or more of the conditions described herein

In another aspect of the disclosure, there is provided methods forstudying P₂Y₆ receptor activity using the agonists of the disclosure,which include the compounds of all formulae disclosed herein, all of theindividual compounds disclosed herein, and all of their prodrugs andsalts. For example, cells in culture may be contacted with one or moreof the compounds provided herein and their impact on P₂Y₆ receptoractivity, as well as cellular function, can be evaluated. Such studiesare useful for evaluating the use of agonists of the disclosure as invitro research tools for evaluating P₂Y₆ receptor activity and itscellular and biochemical impact on different cell types.

In another aspect of the disclosure, there is provided a method ofmodulating P₂Y₆ receptor activity by using one or more of the compoundsdescribed herein, or their prodrugs and/or salts. For example, theinvention provides a method of modulating P₂Y₆ receptor activity in apatient in need thereof by administering to the patient atherapeutically effective amount of one or more of the compoundsdescribed herein, or their prodrugs and/or salts Similarly, theinvention provides a method of modulating P₂Y₆ receptor activity in apatient in need thereof by administering to the patient atherapeutically effective amount of one or more of the compoundsdescribed herein, or their prodrugs and/or salt in a pharmaceuticalcomposition.

In another aspect of the disclosure, there is provided a method fortreating neuronal disorders, including neurodegenerative disorders(e.g., Alzheimer's disease, Parkinson's disease) and traumatic CNSinjury, pain, Down Syndrome (DS), glaucoma and inflammatory conditionsin a subject in need or at risk thereof using a compound describedherein.

In another aspect, the disclosure provides methods for decreasing plaqueburden, improving cognitive function, decreasing or delaying cognitiveimpairment, improving or restoring memory, enhancing synapticplasticity, or improving hippocampal long term potentiation byadministering to a subject in need or at risk thereof a P₂Y₆ receptoragonist. Also provided are methods of enhancing beta amyloid clearance.Subjects in need include subjects having Alzheimer's disease (includingsubjects suspected of having Alzheimer's disease). Additional subjectsin need thereof are subjects having Down Syndrome, and administration ofa P₂Y₆ receptor agonist or a P₂Y₆ receptor-modulating compound is usedto treat Down Syndrome by, for example, improving cognitive function,decreasing cognitive impairment, improving or restoring memory,improving hippocampal long term potentiation, enhancing synapticplasticity, or enhancing clearance of beta amyloid. Further subjects inneed thereof are subjects having Parkinson's disease. Exemplary P₂Y₆receptor agonists are disclosed herein.

In another aspect, the disclosure provides methods for clearing orotherwise decreasing extracellular alpha-synuclein, decreasingintracellular accumulation of alpha-synuclein, and/or decreasing orpreventing the formation of Lewy bodies in a subject in need thereof byadministrating a P₂Y₆ receptor agonist to the subject. In certainembodiments, the subject in need thereof is a subject having Parkinson'sdisease, and administration of a compound of the disclosure provides amethod of treating Parkinson's disease by, for example, improving orpreventing further motor impairment associated with Parkinson's diseaseand/or improving or preventing memory impairment and other symptoms ofneurodegeneration. Without being bound by theory, phagocytosis ofextracellular alpha-synuclein, which may be promoted by the P₂Y₆receptor agonists, decreases extracellular and intracellularaccumulation of alpha-synuclein, as well as Lewy body formation and theresulting neurodegeneration.

In another aspect, the disclosure provides methods for treating glaucomaby administering to a subject in need thereof an effective amount of oneor more of the compounds, salts, or prodrugs disclosed herein. Incertain embodiments, administration of an effective amount of one ormore of the compounds, salts, or prodrugs can decrease intraocularpressure in the subject in need thereof.

In another aspect, the disclosure provides methods for treating aninflammatory condition in a subject in need thereof comprisingadministering to the subject an effective amount of one or more of thecompounds, salts, or prodrugs according to the present disclosure. Incertain embodiments, the disclosure provides a method for reducing theplasma concentration of one or more cytokines in plasma of a subject,such as a subject with an inflammatory condition. Suitable cytokines aredescribed herein. In either case, the disclosure provides numerousexamples of inflammatory conditions which may be treated (e.g., thesubject in need thereof has an inflammatory condition described herein).In certain embodiments, the subject is administered an effective amountof a compound, salt or prodrug of the disclosure. In certainembodiments, the inflammatory condition is not Alzheimer's diseaseand/or the subject being treated does not have, and/or has not beendiagnosed with, and/or is not suspected of having Alzheimer's disease.

In certain embodiments, the disclosure provides a method of treating aninflammatory condition characterized, in whole or in part, by elevatedIL-12 and/or increased IL-12 activity by administering a compound, saltor prodrug of the disclosure. Exemplary conditions are described herein.Similarly the disclosure provides a method of treating an inflammatorycondition characterized, in whole or in part, by elevated levels of oneor more of IL-4, IL-10, or IL-12. Methods of reducing the plasmaconcentration of one or more of any of these cytokines in plasma arealso provided.

In certain embodiments, the disclosure provides methods for treating oneor more of: rheumatoid arthritis, psoriasis, psoriatic arthritis,atherosclerosis, Crohn's disease, ulcerative colitis, irritable bowelsyndrome, or multiple sclerosis.

In certain embodiments, the disclosure provides methods for treating anyof the conditions described herein, such as in a subject at risk fordeveloping the condition, by initiating treatment prior to onset of oneor more symptoms and/or prior to achieving a level of decline at whichstandard of care treatment is typically initiated. In such prophylacticembodiments, the disclosure contemplates, in certain embodiments, thattreatment delays onset of symptoms and/or delays further decline and/orreduces severity of symptoms.

The disclosure contemplates combinations of any of the aspects and/orembodiments described herein. For example, any of the compoundsdescribed herein, such as any of the P₂Y₆ modulating compounds (e.g.,compounds that modulate P₂Y₆ receptor activity) described herein, may beused in the treatment of any of the conditions described herein, such asby administering an effective amount to a subject in need thereof.Similarly, any of the compounds described herein may be provided ascompositions, such as pharmaceutical compositions, and any suchpharmaceutical compositions may be used in the treatment of any of theconditions described herein.

DETAILED DESCRIPTION OF THE FIGURES

FIG. 1 shows two-photon microscopy images of the amyloid plaques labeledwith methoxyX04 in the barrel cortex in living PSAPP mice: (A) image onday 1; (B) magnified view of the portion of the image in the white boxin FIG. 1A, in which the blood plasma was labeled with Rhodaminedextran; (C) magnified view of the portion of the image in the white boxin FIG. 1A, where the arrows indicate dense core plaques; (D) image ofthe same imaging area on day 4, after the injection of UDP; (E)magnified view of the portion of the image in the white box in FIG. 1D,in which the blood plasma was labeled with Rhodamine dextran; and (F)magnified view of the portion of the image in the white box in FIG. 1D,where the arrows indicate dense core plaques.

FIG. 2 shows a quantitative analysis of the number of plaques, plaqueload and size of cross-section of individual plaques in the barrelcortex in PSAPP mice after treatment with UDP or artificial cerebralspinal fluid (ACSF): (A) quantitative analysis of the number of plaques;(B) quantitative analysis of the plaque load; (C) quantitative analysisof the size of cross-section of plaques; (D) UDP treatment reducesplaque load as shown by significant reductions in day 4/day 1 ratios ofplaque load; and (E) UDP treatment reduces number of plaques as shown bysignificant reductions in day 4/day 1 ratios of plaque load.

FIG. 3 shows postmortem immunohistochemistry analysis of the plaque loadin cortex and hippocampus of PSAPP mice after treatment with UDP.Amyloid beta peptide specific antibodies β1-40 and β 1-42 were used inthe immunohistochemistry analysis: (A) immunohistochemistry analysisusing β1-40 on day 1; (B) immunohistochemistry analysis using β1-40 onday 4, after treatment with UDP; (C) immunohistochemistry analysis usingβ1-42 on day 1; and (D) immunohistochemistry analysis using β1-42 on day4, after treatment with UDP.

FIG. 4 shows quantification of plaque load (%) in the cortex andhippocampus of the PSAPP mice after treatment with UDP or ACSF. Amyloidbeta peptide specific antibodies β1-40 and β1-42 were used in thequantification. (A) plaque load (%) in cortex using β1-40 staining; (B)plaque load (%) in hippocampus using β1-40 staining; (C) plaque load (%)in cortex using β1-42 staining; (D) plaque load (%) in hippocampus usingβ1-42 staining; (E) UDP treatment decreased soluble Aβ40 level detectedwith ELISA; and (F) UDP treatment decreased soluble Aβ42 level detectedwith ELISA.

FIG. 5 shows a postmortem immunohistochemistry analysis of the plaqueload in cortex and hippocampus of PSAPP mice after intraperitoneal(i.p.) injection of 3-phenacyl-UDP for 2, 4 and 6 consecutive days.Amyloid beta specific antibody β1-40 was used in the analysis. (A)immunohistochemistry analysis using β1-40 without 3-phenacyl-UDPtreatment; (B) immunohistochemistry analysis using β1-40 afterintraperitoneal injection of 3-phenacyl-UDP for 2 consecutive days; (C)immunohistochemistry analysis using β1-40 after intraperitonealinjection of 3-phenacyl-UDP for 4 consecutive days; and (D)immunohistochemistry analysis using β1-40 after intraperitonealinjection of 3-phenacyl-UDP for 6 consecutive days.

FIG. 6 shows quantification of plaque load (%) in cortex (Cx) andhippocampus (Hp) of the PSAPP mice after treatment with 3-phenacyl-UDPor vehicle control for 2, 4, 6 consecutive days and for 6 days+2 weeks.The vehicle controls used for intracerebroventricular (icv) andIntraperitoneal (ip) administration of compounds were ACSF and saline,respectively. Amyloid beta peptide specific antibody β1-40 was used inquantification. (A) Plaque load (%) in cortex using β1-40 staining; (B)plaque load (%) in hippocampus using β1-40 staining; (C) Aβ40 plaqueload (%) in hippocampus after one week of daily treatment with3-phenacyl-UDP (PSB0474) at three doses; (D) Aβ42 plaque load (%) inhippocampus after one week of daily treatment with 3-phenacyl-UDP(PSB0474) at three doses; (E) Aβ40 plaque load (%) in cortex after oneweek of daily treatment with 3-phenacyl-UDP (PSB0474) at three doses;and (F) Aβ42 plaque load (%) in cortex after one week of daily treatmentwith 3-phenacyl-UDP (PSB0474) at three doses.

FIG. 7 shows freezing behavior (freezing %) of PASPP mice in fearconditioning studies after treatment with ACSF or UDP: (A) freezingbehavior (freezing %) of PASPP mice 5 minutes following treatment withACSF and UDP; (B) analysis of total freezing percentage of PSAPP micetreated with ACSF or UDP; and (C) using the contextual fear conditioningtest PSAPP mice treated with ACSF (white bar) showed significantly lessfreezing time compared to the age-matched wildtype (line bar),suggesting the memory deficits in PS1/APP; UDP-treatment 3 days prior tothe test significantly improved the freezing behavior (black bar)compared to ACSF treatment.

FIG. 8 shows hippocampal long-term potentiation (LTP) recorded as fieldexcitatory postsynaptic potential (fEPSP) % in PSAPP mice, withhigh-frequency stimulation (HFS), 100 pulses at 100 Hz, four times in20-second intervals: (A) depressed LTP (fEPSP %) at the schaffercollateral synapse within the CA1 area of the hippocampus in aged PSAPPmice (PSAPP+/+), as compared to littermates (PSAPP−/−); (B) increasedLTP (fEPSP %) in PSAPP mice after treatment with UDP or ACSF; (C)analysis of the last 15 min potentiation, as fEPSP slope (%), in PSAPPmice.

FIG. 9 shows freezing behavior (as freezing %) of PASPP mice in fearconditioning studies after treatment with 3-phenacyl-UDP (PSB0474). (A)freezing behavior (freezing %) of control littermates (PSAPP−/−), andPASPP mice 5 minutes following treatment with saline vehicle control orwith 3-phenacyl-UDP (PSB0474) at two different dosages, i.e. 1 μg/ml and1 mg/ml; (B) analysis of total freezing percentage of PSAPP mice; and(C) using the contextual fear conditioning test PSAPP mice treated withACSF (white bar) showed significantly less freezing time compared to theage-matched wildtype (line bar), demonstrating the memory deficits inPS1/APP; one week treatment with 1 μg/kg 3-phenacyl-UDP (PSB0474) (greybar) rescued the memory deficit as compared to the vehicle treatment(white bar).

FIG. 10 shows dose-response activation of the P₂Y₆ receptor usingcompounds of the present disclosure, where compounds were tested foractivation of P₂Y₆ receptor by measuring receptor induced Ca²⁺ changeswith the fluorescent Ca²⁺ indicator fluo-4: (A) dose-response activationof the P₂Y₆ receptor using the sodium salt of the diphosphate derivativeof compound 6; (B) dose-response activation of the P₂Y₆ receptor usingthe sodium salt of the diphosphate derivative of compound 3; (C)dose-response activation of the P₂Y₆ receptor using the sodium salt ofthe diphosphate derivative of compound 4; (D) dose-response activationof the using the sodium salt of the diphosphate derivative of compound1; (E) dose-response activation of the P₂Y₆ receptor using the sodiumsalt of the diphosphate derivative of compound 5; (F) dose-responseactivation of the P₂Y₆ receptor using the sodium salt of the diphosphatederivative of compound 32; (G) dose-response activation of the P₂Y₆receptor using the sodium salt of the diphosphate derivative of compound33; (H) dose-response activation of the P₂Y₆ receptor using the sodiumsalt of the diphosphate derivative of compound 34; (I) dose-responseactivation of the P₂Y₆ receptor using the sodium salt of the diphosphatederivative of compound 35; (J) dose-response activation of the P₂Y₆receptor using the sodium salt of the diphosphate derivative of compound36; and (K) dose-response activation of the P₂Y₆ receptor using thesodium salt of the diphosphate derivative of compound 37.

FIG. 11 shows freezing behavior (freezing %) of PASPP mice in fearconditioning studies after treatment with vehicle control or thediphosphate derivative of compound 5: using the contextual fearconditioning test PSAPP mice treated with vehicle control (black bar)showed significantly less freezing time compared to the age-matchedwildtype (white bar), suggesting the memory deficits in PSAPP;administration of the diphosphate derivative of compound 5 prior to thetest significantly improved the freezing behavior (line bar) compared tothe control treatment indicating that the diphosphate derivative ofcompound 5 restores memory.

FIG. 12 shows plaque load in cortex (Cx) and hippocampus (Hp) of thePSAPP mice after treatment with the diphosphate derivative of compound 5or vehicle control. (A) Aβ plaque load (%) in cortex after treatmentwith the diphosphate derivative of compound 5 or vehicle control; (B) Aβplaque load (%) in hippocampus after treatment with the diphosphatederivative of compound 5 or vehicle control; and (C) postmortemimmunohistochemistry analysis of the Aβ42 plaque load in cortex andhippocampus of PSAPP mice after treatment with the diphosphatederivative of compound 5 or vehicle control. Amyloid beta specificantibody β1-42 was used in the analysis.

FIG. 13 shows quantification of plaque load (%) in the cortex of thePSAPP mice after treatment with nucleoside compound 5. Amyloid betapeptide specific antibodies β1-40 and β1-42 were used in thequantification. (A) plaque load (%) in cortex using β1-40 staining; (B)plaque load (%) in cortex using β1-42 staining.

FIG. 14 summarizes plasma cytokine levels (pg/ml) in wildtype and PSAPPmice treated for 7 days (i.p.) with vehicle or the diphosphatederivative of compound 5 (1 μg/kg). Plasma cytokine levels are measuredin pg/ml. The x-axis of the graph shows the various cytokines examinedand the y-axis represents concentration. For each cytokine shown alongthe x-axis, the graph provides four bars indicative of the results forthe various treatment groups which are, from left to right: untreatedwildtype animals, wildtype animals treated with the diphosphatederivative compound 5, untreated PSAPP animals, and PSAPP animalstreated with the diphosphate derivative compound 5.

FIG. 15 summarizes results of a fear conditioning assay. PSAPP mice weretreated daily for 100 days with vehicle or compound 5 (10 μg/kg; denotedon the figure as “compound”). Compound or vehicle was administeredintraperitoneally (i.p.). The mice were then assessed in a fearconditioning task for memory formation.

FIG. 16 summarizes results illustrating decrease in plaque burdenfollowing 100 days of treatment with compound 5.

FIG. 17 summarizes plasma cytokine levels (pg/ml) in PSAPP mice treateddaily for 100 days with vehicle or compound 5 (10 μg/kg). Compound orvehicle was administered intraperitoneally (i.p.). Concentration ofplasma cytokine levels are measured in pg/ml, as represented on they-axis. For each cytokine examined, plasma levels of vehicle treated orcompound treated animals are shown, as represented on the x-axis. Datafor the following cytokines are presented: IL-4, IL-9, IL-5, IL-10,IL-6, IL-12, and IL-7. For IL-12, levels of just the p40 subunit (p40),as well as levels of the heterodimeric cytokine (p70) are assayed.

FIG. 18 summarizes plasma cytokine levels (pg/ml) in PSAPP mice treatedfor 100 days with vehicle or compound 5 (10 μg/kg). Compound or vehiclewas administered intraperitoneally (i.p.). Concentration of plasmacytokine levels are measured in pg/ml, as represented on the y-axis. Foreach cytokine examined, plasma levels of vehicle treated or compoundtreated animals are shown, as represented on the x-axis. Data for thefollowing cytokines are presented: eotaxin, IL-1α, G-CSF, IL-1β, GM-CSF,IL-2, IFN-r, IL-3.

FIG. 19 summarizes plasma cytokine levels (pg/ml) in PSAPP mice treatedfor 100 days with vehicle or compound 5 (10 μg/kg). Compound or vehiclewas administered intraperitoneally (i.p.). Concentration of plasmacytokine levels are measured in pg/ml, as represented on the y-axis. Foreach cytokine examined, plasma levels of vehicle treated or compoundtreated animals are shown, as represented on the x-axis. Data for thefollowing cytokines are presented: IL-13, KC, IL-15, LIF, IL-17, LIX,IP-10, and MCP-1.

FIG. 20 summarizes plasma cytokine levels (pg/ml) in PSAPP mice treatedfor 100 days with vehicle or compound 5 (10 μg/kg). Compound or vehiclewas administered intraperitoneally (i.p.). Concentration of plasmacytokine levels are measured in pg/ml, as represented on the y-axis. Foreach cytokine examined, plasma levels of vehicle treated or compoundtreated animals are shown, as represented on the x-axis. Data for thefollowing cytokines are presented: M-CSF, MIP2, MIG, RANTES, MIP-1a, andMIP-1b, and TNFα.

FIG. 21 summarizes plasma cytokine levels (pg/ml) in PSAPP mice treatedfor 100 days with vehicle or compound 5 (10 μg/kg), with treatmentbeginning at approximately 3 months of age. Compound or vehicle wasadministered intraperitoneally (i.p). Concentration of plasma cytokinelevels are measured in pg/ml, as represented on the y-axis. For eachcytokine examined, plasma levels of vehicle treated or compound treatedanimals are shown, as represented on the x-axis. Data for the followingcytokines are presented: IL-12, IL-13, IL-17, IL-10, MIP-1a, MIP-1b,IL-2, and IL-4. * denotes p<0.05; ** denotes p<0.01.

FIG. 22 summarizes additional data obtained as part of the same study asin FIG. 21 in which PSAPP mice were treated for 100 days with vehicle orcompound 5, with treatment beginning at approximately 3 months of age.Plaque load was evaluated in the cortex (FIG. 22A) or hippocampus (FIG.22B). The mice were also evaluated in a fear conditioning test of memory(FIG. 22C). In FIG. 22A, *** denotes p<0.01. In FIGS. 22B and 22C, *denotes p<0.02.

FIG. 23 summarizes plasma cytokine levels (pg/ml) in PSAPP mice treatedfor 7 days with vehicle or compound 5 (10 μg/kg), with treatmentbeginning at approximately 6 months of age after onset of symptoms inthese PSAPP mice. Compound or vehicle was administered intraperitoneally(i.p). Concentration of plasma cytokine levels are measured in pg/ml, asrepresented on the y-axis. For each cytokine examined, plasma levels ofvehicle treated or compound treated animals are shown, as represented onthe x-axis. Data for the following cytokines are presented: IL-12,IL-13, IL-17, IL-10, MIP-1a, and TNFalpha. * denotes p<0.05; ** denotesp<0.005; *** denotes p<0.0005.

FIG. 24 summarizes additional data obtained as part of the same study asin FIG. 23 in which PSAPP mice were treated for 7 days with vehicle orcompound 5 (10 μg/kg), with treatment beginning at approximately 6months of age after onset of symptoms in these PSAPP mice. Plaque loadwas evaluated in the cortex (FIG. 24A, top panel) and hippocampus (FIG.24A, bottom panel). In addition to plaque load, plaque size wasevaluated (FIG. 24B). * denotes p<0.05.

FIG. 25 summarize data evaluating cytokine release from human THP-1cells treated in vitro with vehicle or the P₂Y₆ antagonist MRS 2578 (CASNo. 711019-86-2). Concentration of cytokine secreted into the culturemedia is measured in pg/ml, as represented on the y-axis. For eachcytokine examined, levels of cytokine released into the culture mediafollowing treatment with vehicle or MRS 2578, as represented on thex-axis, are shown. Results for the following cytokines are depicted:fractalkine (FIG. 25A), PDGF-BB (FIG. 25B), and IL-7 (FIG. 25C).Treatment with MRS 2578 increased release of each of these threecytokines from human THP-1 cells, relative to treatment with vehiclecontrol. Increased release is assayed by measuring concentration of thecytokine in the cell culture supernatant following treatment. * denotesp<0.05; ** denotes p<0.02; *** denotes p<0.002.

FIG. 26 summarizes data evaluating concentration of cytokine (IL-7)released from human THP-1 cells treated in vitro with vehicle, compound5, the P₂Y₆ antagonist MRS 2578, or both compound 5 and MRS 2578.Concentration of cytokine secreted into the culture media is measured inpg/ml, as represented on the y-axis. Treatment with MRS 2578 increasedrelease of IL-7 from human THP-1 cells. Treatment with compound 5decreased release of IL-7 from human THP-1 cells, and this effect wasabrogated by co-treatment with MRS 2578. * denotes p<0.05.

DETAILED DESCRIPTION OF THE INVENTION

A. Definitions

Unless otherwise defined herein, scientific and technical terms used inthis application shall have the meanings that are commonly understood bythose of ordinary skill in the art. Generally, nomenclature used inconnection with, and techniques of, chemistry, cell and tissue culture,molecular biology, cell and cancer biology, neurobiology,neurochemistry, virology, immunology, microbiology, pharmacology,genetics and protein and nucleic acid chemistry, described herein, arethose well known and commonly used in the art.

The methods and techniques of the present disclosure are generallyperformed, unless otherwise indicated, according to conventional methodswell known in the art and as described in various general and morespecific references that are cited and discussed throughout thisspecification. See, e.g. “Principles of Neural Science”, McGraw-HillMedical, New York, N.Y. (2000); Motulsky, “Intuitive Biostatistics”,Oxford University Press, Inc. (1995); Lodish et al., “Molecular CellBiology, 4th ed.”, W. H. Freeman & Co., New York (2000); Griffiths etal., “Introduction to Genetic Analysis, 7th ed.”, W. H. Freeman & Co.,N.Y. (1999); and Gilbert et al., “Developmental Biology, 6th ed.”,Sinauer Associates, Inc., Sunderland, Mass. (2000).

Chemistry terms used herein are used according to conventional usage inthe art, as exemplified by “The McGraw-Hill Dictionary of ChemicalTerms”, Parker S., Ed., McGraw-Hill, San Francisco, Calif. (1985).

All of the above, and any other publications, patents and publishedpatent applications referred to in this application are specificallyincorporated by reference herein. In case of conflict, the presentspecification, including its specific definitions, will control.

The term “agent” is used herein to denote a chemical compound (such asan organic or inorganic compound, a mixture of chemical compounds), abiological macromolecule (such as a nucleic acid, an antibody, includingparts thereof as well as humanized, chimeric and human antibodies andmonoclonal antibodies, a protein or portion thereof, e.g., a peptide, alipid, a carbohydrate), or an extract made from biological materialssuch as bacteria, plants, fungi, or animal (particularly mammalian)cells or tissues. Agents include, for example, agents that are knownwith respect to structure, and those that are not known with respect tostructure. The P₂Y₆ receptor-modulating activity (such as direct orindirect agonist activity) of such agents may render them suitable as“therapeutic agents” in the methods and compositions of this disclosure.

A “patient,” “subject,” or “individual” are used interchangeably andrefer to either a human or a non-human animal. These terms includemammals, such as humans, primates, livestock animals (including bovines,porcines, etc.), companion animals (e.g., canines, felines, etc.) androdents (e.g., mice and rats).

“Treating” a condition or patient refers to taking steps to obtainbeneficial or desired results, including clinical results. Beneficial ordesired clinical results include, but are not limited to, alleviation,amelioration, or slowing the progression, of one or more symptomsassociated with a neuronal disorder, including neurodegeneration andtraumatic brain injury, as well as pain. In certain embodiments,treatment may be prophylactic. Exemplary beneficial clinical results aredescribed herein.

“Administering” or “administration of” a substance, a compound or anagent to a subject can be carried out using one of a variety of methodsknown to those skilled in the art. For example, a compound or an agentcan be administered, intravenously, arterially, intradermally,intramuscularly, intraperitoneally, subcutaneously, ocularly,sublingually, orally (by ingestion), intranasally (by inhalation),intraspinally, intracerebrally, and transdermally (by absorption, e.g.,through a skin duct). A compound or agent can also appropriately beintroduced by rechargeable or biodegradable polymeric devices or otherdevices, e.g., patches and pumps, or formulations, which provide for theextended, slow or controlled release of the compound or agent.Administering can also be performed, for example, once, a plurality oftimes, and/or over one or more extended periods. In some aspects, theadministration includes both direct administration, includingself-administration, and indirect administration, including the act ofprescribing a drug. For example, as used herein, a physician whoinstructs a patient to self-administer a drug, or to have the drugadministered by another and/or who provides a patient with aprescription for a drug is administering the drug to the patient.

Appropriate methods of administering a substance, a compound or an agentto a subject will also depend, for example, on the age of the subject,whether the subject is active or inactive at the time of administering,whether the subject is cognitively impaired at the time ofadministering, the extent of the impairment, and the chemical andbiological properties of the compound or agent (e.g. solubility,digestibility, bioavailability, stability and toxicity). In someembodiments, a compound or an agent is administered orally, e.g., to asubject by ingestion. In some embodiments, the orally administeredcompound or agent is in an extended release or slow release formulation,or administered using a device for such slow or extended release.

A “therapeutically effective amount” or a “therapeutically effectivedose” of a drug or agent is an amount of a drug or an agent that, whenadministered to a subject will have the intended therapeutic effect. Thefull therapeutic effect does not necessarily occur by administration ofone dose, and may occur only after administration of a series of doses.Thus, a therapeutically effective amount may be administered in one ormore administrations. The precise effective amount needed for a subjectwill depend upon, for example, the subject's size, health and age, thenature and extent of cognitive impairment or other symptoms of thecondition being treated, such as neurodegeneration (such as Alzheimer'sdisease), pain and traumatic brain injury, the therapeutics orcombination of therapeutics selected for administration, and the mode ofadministration. The skilled worker can readily determine the effectiveamount for a given situation by routine experimentation.

“Ligand” as used herein refers to any molecule that is capable ofspecifically binding to another molecule, such as the P₂Y₆ receptor. Theterm “ligand” includes both agonists and antagonists. “Agonist” means anagent which, when interacting, either directly or indirectly, with abiologically active molecule (e.g. an enzyme or a receptor) causes anincrease in the biological activity thereof. “Antagonist” means an agentwhich, when interacting, either directly or indirectly, with abiologically active molecule(s) (e.g. an enzyme or a receptor) causes adecrease in the biological activity thereof. In certain embodiments, thecompounds of the present disclosure modulate P₂Y₆ receptor activity,either directly or indirectly. In certain embodiments, the compoundsagonize P₂Y₆ receptor activity, for example, directly, for example, bydirect interaction with the P₂Y₆ receptor, or indirectly, for example,via a metabolite that interacts with the P₂Y₆ receptor. In certainembodiments, the compounds of the disclosure (such as compounds offormulae I (including compounds of formula I-A and formula I-B) and IIand pharmaceutically acceptable salts and prodrugs thereof, as well asthe individual compounds disclosed herein) are used, directly orindirectly, as P₂Y₆ receptor agonists or P₂Y₆ receptor-modulatingcompounds, and may be used in any of the in vitro and/or in vivo methodsdisclosed herein. In certain embodiments, compounds disclosed herein arethemselves P₂Y₆ receptor-modulating compounds, and the disclosureencompasses these compounds as well as their salts and/or prodrugs asagonists of the disclosure. Other compounds, salts, and prodrugsdescribed herein are not active themselves, but are converted in vivo tocompounds that are active P₂Y₆ receptor-modulating compounds. Thedisclosure contemplates that all such compounds, salts, or prodrugs ofthe disclosure, whether active themselves or are converted into activecompounds in vivo, may be used to treat any of conditions describedherein.

The term “aliphatic” as used herein means a straight chained or branchedalkyl, alkenyl or alkynyl. It is understood that alkenyl or alkynylembodiments need at least two carbon atoms in the aliphatic chain.Aliphatic groups typically contains from 1 (or 2) to 12 carbons, such asfrom 1 (or 2) to 4 carbons.

The term “aryl” as used herein means a monocyclic or bicycliccarbocyclic aromatic ring system. Phenyl is an example of a monocyclicaromatic ring system. Bicyclic aromatic ring systems include systemswherein both rings are aromatic, e.g., naphthyl, and systems whereinonly one of the two rings is aromatic, e.g., tetralin.

The term “heterocyclic” as used herein means a monocyclic or bicyclicnon-aromatic ring system having 1 to 3 heteroatom or heteroatom groupsin each ring selected from O, N, NH, S, SO, or SO₂ in a chemicallystable arrangement. In a bicyclic non-aromatic ring system embodiment of“heterocyclyl”, one or both rings may contain said heteroatom orheteroatom groups. In another heterocyclic ring system embodiment, anon-aromatic heterocyclic ring may optionally be fused to an aromaticcarbocycle.

Examples of heterocyclic rings include 3-1H-benzimidazol-2-one,3-(1-alkyl)-benzimidazol-2-one, 2-tetrahydrofuranyl,3-tetrahydrofuranyl, 2-tetrahydrothiophenyl, 3-tetrahydrothiophenyl,2-morpholino, 3-morpholino, 4-morpholino, 2-thiomorpholino,3-thiomorpholino, 4-thiomorpholino, 1-pyrrolidinyl, 2-pyrrolidinyl,3-pyrrolidinyl, 1-tetrahydropiperazinyl, 2-tetrahydropiperazinyl,3-tetrahydropiperazinyl, 1-piperidinyl, 2-piperidinyl, 3-piperidinyl,1-pyrazolinyl, 3-pyrazolinyl, 4-pyrazolinyl, 5-pyrazolinyl,1-piperidinyl, 2-piperidinyl, 3-piperidinyl, 4-piperidinyl,2-thiazolidinyl, 3-thiazolidinyl, 4-thiazolidinyl, 1-imidazolidinyl,2-imidazolidinyl, 4-imidazolidinyl, 5-imidazolidinyl, indolinyl,tetrahydroquinolinyl, tetrahydroisoquinolinyl, benzothiolane,benzodithiane, and 1,3-dihydro-imidazol-2-one.

The term “heteroaryl” as used herein means a monocyclic or bicyclicaromatic ring system having 1 to 3 heteroatom or heteroatom groups ineach ring selected from O, N, NH or S in a chemically stablearrangement. In such a bicyclic aromatic ring system embodiment of“heteroaryl” both rings may be aromatic; and one or both rings maycontain said heteroatom or heteroatom groups.

Examples of heteroaryl rings include 2-furanyl, 3-furanyl, N-imidazolyl,2-imidazolyl, 4-imidazolyl, 5-imidazolyl, benzimidazolyl, 3-isoxazolyl,4-isoxazolyl, 5-isoxazolyl, 2-oxazolyl, 4-oxazolyl, 5-oxazolyl,N-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl, 2-pyridyl, 3-pyridyl, 4-pyridyl,2-pyrimidinyl, 4-pyrimidinyl, 5-pyrimidinyl, pyridazinyl (e.g.,3-pyridazinyl), 2-thiazolyl, 4-thiazolyl, 5-thiazolyl, tetrazolyl (e.g.,5-tetrazolyl), triazolyl (e.g., 2-triazolyl and 5-triazolyl), 2-thienyl,3-thienyl, benzofuryl, benzothiophenyl, indolyl (e.g., 2-indolyl),pyrazolyl (e.g., 2-pyrazolyl), isothiazolyl, 1,2,3-oxadiazolyl,1,2,5-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,3-triazolyl,1,2,3-thiadiazolyl, 1,3,4-thiadiazolyl, 1,2,5-thiadiazolyl, purinyl,pyrazinyl, 1,3,5-triazinyl, quinolinyl (e.g., 2-quinolinyl,3-quinolinyl, 4-quinolinyl), and isoquinolinyl (e.g., 1-isoquinolinyl,3-isoquinolinyl, or 4-isoquinolinyl).

The term “cycloalkyl or cycloalkenyl” refers to a monocyclic or fused orbridged bicyclic carbocyclic ring system that is not aromatic.Cycloalkenyl rings have one or more units of unsaturation. Exemplarycycloalkyl or cycloalkenyl groups include cyclopropyl, cyclobutyl,cyclopentyl, cyclohexyl, cyclohexenyl, cycloheptyl, cycloheptenyl,norbornyl, adamantyl and decalinyl.

As used herein, the carbon atom designations may have the indicatedinteger and any intervening integer. For example, the number of carbonatoms in a (C1-C4)-alkyl group is 1, 2, 3, or 4. It should be understoodthat these designation refer to the total number of atoms in theappropriate group. For example, in a (C3-C10)-heterocyclyl the totalnumber of carbon atoms and heteroatoms is 3 (as in aziridine), 4, 5, 6(as in morpholine), 7, 8, 9, or 10.

“Pharmaceutically acceptable salt” or “salt” is used herein to refer toan agent or a compound according to the disclosure that is atherapeutically active, non-toxic base and acid salt form of thecompounds. The acid addition salt form of a compound that occurs in itsfree form as a base can be obtained by treating said free base form withan appropriate acid such as an inorganic acid, for example, a hydrohalicsuch as hydrochloric or hydrobromic, sulfuric, nitric, phosphoric andthe like; or an organic acid, such as, for example, acetic,hydroxyacetic, propanoic, lactic, pyruvic, malonic, succinic, maleic,fumaric, malic, tartaric, citric, methanesulfonic, ethanesulfonic,benzenesulfonic, p-toluenesulfonic, cyclic, salicylic, p-aminosalicylic,pamoic and the like. See, e.g., WO 01/062726.

Compounds containing acidic protons may be converted into theirtherapeutically active, non-toxic base addition salt form, e. g. metalor amine salts, by treatment with appropriate organic and inorganicbases. Appropriate base salt forms include, for example, ammonium salts,alkali and earth alkaline metal salts, e. g., lithium, sodium,potassium, magnesium, calcium salts and the like, salts with organicbases, e. g. N-methyl-D-glucamine, hydrabamine salts, and salts withamino acids such as, for example, arginine, lysine and the like.Conversely, said salt forms can be converted into the free forms bytreatment with an appropriate base or acid. Compounds and their saltscan be in the form of a solvate, which is included within the scope ofthe present disclosure. Such solvates include for example hydrates,alcoholates and the like. See, e.g., WO 01/062726.

Many of the compounds useful in the methods and compositions of thisdisclosure have at least one stereogenic center in their structure. Thisstereogenic center may be present in a R or a S configuration, said Rand S notation is used in correspondence with the rules described inPure Appl. Chem. (1976), 45, 11-30. The disclosure also relates to allstereoisomeric forms such as enantiomeric and diastereoisomeric forms ofthe compounds or mixtures thereof (including all possible mixtures ofstereoisomers). See, e.g., WO 01/062726.

Furthermore, certain compounds which contain alkenyl groups may exist asZ (zusammen) or E (entgegen) isomers. In each instance, the disclosureincludes both mixture and separate individual isomers. Multiplesubstituents on a piperidinyl or the azepanyl ring can also stand ineither cis or trans relationship to each other with respect to the planeof the piperidinyl or the azepanyl ring. Some of the compounds may alsoexist in tautomeric forms. Such forms, although not explicitly indicatedin the formulae described herein, are intended to be included within thescope of the present disclosure. With respect to the methods andcompositions of the present disclosure, reference to a compound orcompounds is intended to encompass that compound in each of its possibleisomeric forms and mixtures thereof unless the particular isomeric formis referred to specifically. See, e.g., WO 01/062726.

“Prodrug” or “pharmaceutically acceptable prodrug” refers to a compoundthat is metabolized, for example hydrolyzed or oxidized, in the hostafter administration to form the compound of the present disclosure(e.g., compounds of formula I or II). Typical examples of prodrugsinclude compounds that have biologically labile or cleavable(protecting) groups on a functional moiety of the active compound.Prodrugs include compounds that can be oxidized, reduced, aminated,deaminated, hydroxylated, dehydroxylated, hydrolyzed, dehydrolyzed,alkylated, dealkylated, acylated, deacylated, phosphorylated, ordephosphorylated to produce the active compound. Examples of prodrugsusing ester or phosphoramidate as biologically labile or cleavable(protecting) groups are disclosed in U.S. Pat. Nos. 6,875,751,7,585,851, and 7,964,580, the disclosures of which are incorporatedherein by reference. The prodrugs of this disclosure are metabolized toproduce a compound of formula I or II, which are agonists or modulatorsof the P₂Y₆ receptor.

The disclosure further provides pharmaceutical compositions comprisingone or more compounds of the disclosure together with a pharmaceuticallyacceptable carrier or excipient. Compounds or pharmaceuticalcompositions of the disclosure may be used in vitro or in vivo.

B. Uridine Nucleoside Derivatives and Compositions

The present disclosure provides a compound of formula I:

-   or a prodrug or salt thereof, wherein:-   A is a 3- to 10-membered aromatic or non-aromatic ring having up to    5 heteroatoms independently selected from N, O, S, SO, or SO₂,    wherein the aromatic or non-aromatic ring is independently and    optionally substituted with one or more R⁷;-   X is selected from:    -   —H, —C(O)R⁵, —C(O)OR⁵, and —P(O)(OR⁵)₂, preferably X is —H,        —C(O)R⁵, or —C(O)OR⁵;-   Y is a bond or a (C1-C5)-aliphatic group independently and    optionally substituted with one or more R⁴;-   Z and W are each independently selected from ═O, ═S, ═N(R⁵), and    ═NOR⁵;-   R¹ is selected from:    -   —H, halogen, —OR⁵, —CN, —CF₃, —OCF₃ and a (C1-C6)-aliphatic        group optionally substituted with one or more R⁷;-   R² and R³ are each independently selected from —OR⁵, —SR⁵, —NR⁵R⁶,    —OC(O)R⁵, —OC(O)NR⁵R⁶, and —OC(O)OR⁵; preferably, R² and R³ are each    independently selected from —OR⁵, —SR⁵, —NR⁵R⁶ and —OC(O)R⁵;-   each occurrence of R⁴ is independently selected from:    -   halogen, —OR⁵, —NO₂, —CN, —CF₃, —OCF₃, —R⁵, 1,2-methylenedioxy,        1,2-ethylenedioxy, —N(R⁵)₂, —SR⁵, —SOR⁵, —SO₂R⁵, —SO₂N(R⁵)₂,        —SO₃R⁵, —C(O)R⁵, —C(O)C(O)R⁵, —C(O)CH₂C(O)R⁵, —C(S)R⁵, —C(S)OR⁵,        —C(O)OR⁵, —C(O)C(O)OR⁵, —C(O)C(O)N(R⁵)₂, —OC(O)R⁵, —C(O)N(R⁵)₂,        —OC(O)N(R⁵)₂, —C(S)N(R⁵)₂, —(CH₂)₀₋₂NHC(O)R⁵, —N(R⁵)N(R⁵)COR⁵,        —N(R⁵)N(R⁵)C(O)OR⁵, —N(R⁵)N(R⁵)CON(R⁵)₂, —N(R⁵)SO₂R⁵,        —N(R⁵)SO₂N(R⁵)₂, —N(R⁵)C(O)OR⁵, —N(R⁵)C(O)R⁵, —N(R⁵)C(S)R⁵,        —N(R⁵)C(O)N(R⁵)₂, —N(R⁵)C(S)N(R⁵)₂, —N(COR⁵)COR⁵, —N(OR⁵)R⁵,        —C(═NH)N(R⁵)₂, —C(O)N(OR⁵)R⁵, —C(═NOR⁵)R⁵, —OP(O)(OR⁵)₂,        —P(O)(R⁵)₂, —P(O)(OR⁵)₂, or —P(O)(H)(OR⁵);-   each occurrence of R⁵ is independently selected from:    -   H—,    -   (C1-C12)-aliphatic-,    -   (C3-C10)-cycloalkyl- or -cycloalkenyl-,    -   [(C3-C10)-cycloalkyl or -cycloalkenyl]-(C1-C12)-aliphatic-,    -   (C6-C10)-aryl-,    -   (C6-C10)-aryl-(C1-C12)aliphatic-,    -   (C3-C10)-heterocyclyl-,    -   (C6-C10)-heterocyclyl-(C1-C12)aliphatic-,    -   (C5-C10)-heteroaryl-, and    -   (C5-C10)-heteroaryl-(C1-C12)-aliphatic-;    -   wherein two R⁵ groups bound to the same atom optionally form a        3- to 10-membered aromatic or non-aromatic ring having up to 3        heteroatoms independently selected from N, O, S, SO, or SO₂,        wherein said ring is optionally fused to a (C6-C10)aryl,        (C5-C10)heteroaryl, (C3-C10)cycloalkyl, or a        (C3-C10)heterocyclyl; and    -   wherein each R⁵ group is independently and optionally        substituted with one or more R⁷;-   R⁶ is selected from:    -   —R⁵, —C(O)R⁵, —C(O)OR⁵, —C(O)N(R⁵)₂ and —S(O)₂R⁵;-   each occurrence of R⁷ is independently selected from:    -   halogen, —OR⁸, —NO₂, —CN, —CF₃, —OCF₃, —R⁸, oxo, thioxo,        1,2-methylenedioxy, 1,2-ethylenedioxy, —N(R⁸)₂, —SR⁸, —SOR⁸,        —SO₂R⁸, —SO₂N(R⁸)₂, —SO₃R⁸, —C(O)R⁸, —C(O)C(O)R⁸,        —C(O)CH₂C(O)R⁸, —C(S)R⁸, —C(S)OR⁸, —C(O)OR⁸, —C(O)C(O)OR⁸,        —C(O)C(O)N(R⁸)₂, —OC(O)R⁸, —C(O)N(R⁸)₂, —OC(O)N(R⁸)₂,        —C(S)N(R⁸)₂, —(CH₂)₀₋₂NHC(O)R⁸, —N(R⁸)N(R⁸)COR⁸,        —N(R⁸)N(R⁸)C(O)OR⁸, —N(R⁸)N(R⁸)CON(R⁸)₂, —N(R⁸)SO₂R⁸,        —N(R⁸)SO₂N(R⁸)₂, —N(R⁸)C(O)OR⁸, —N(R⁸)C(O)R⁸, —N(R⁸)C(S)R⁸,        —N(R⁸)C(O)N(R⁸)₂, —N(R⁸)C(S)N(R⁸)₂, —N(COR⁸)COR⁸, —N(OR⁸)R⁸,        —C(═NH)N(R⁸)₂, —C(O)N(OR⁸)R⁸, —C(═NOR⁸)R⁸, —OP(O)(OR⁸)₂,        —P(O)(R⁸)₂, —P(O)(OR⁸)₂, or —P(O)(H)(OR⁸);-   each occurrence of R⁸ is independently selected from:    -   H— and (C1-C6)-aliphatic-.

In some embodiments of compound of formula I, each occurrence of R⁴ isindependently selected from:

-   -   halogen, —OR⁵, —NO₂, —CN, —CF₃, —OCF₃, —R⁵, —N(R⁵)₂, —SR⁵,        —SOR⁵, —SO₂R⁵, —SO₂N(R⁵)₂, —SO₃R⁵, —C(O)R⁵, —C(O)C(O)R⁵,        —C(O)CH₂C(O)R⁵, —C(S)R⁵, —C(S)OR⁵, —C(O)OR⁵, —C(O)C(O)OR⁵,        —C(O)C(O)N(R⁵)₂, —OC(O)R⁵, —C(O)N(R⁵)₂, —OC(O)N(R⁵)₂,        —C(S)N(R⁵)₂, —(CH₂)₀₋₂NHC(O)R⁵, —N(R⁵)N(R⁵)COR⁵,        —N(R⁵)N(R⁵)C(O)OR⁵, —N(R⁵)N(R⁵)CON(R⁵)₂, —N(R⁵)SO₂R⁵,        —N(R⁵)SO₂N(R⁵)₂, —N(R⁵)C(O)OR⁵, —N(R⁵)C(O)R⁵, —N(R⁵)C(S)R⁵,        —N(R⁵)C(O)N(R⁵)₂, —N(R⁵)C(S)N(R⁵)₂, —N(COR⁵)COR⁵, —N(OR⁵)R⁵,        —C(═NH)N(R⁵)₂, —C(O)N(OR⁵)R⁵, —C(═NOR⁵)R⁵, —OP(O)(OR⁵)₂,        —P(O)(R⁵)₂, —P(O)(OR⁵)₂, or —P(O)(H)(OR⁵).

In certain embodiments, each occurrence of R⁷ is independently selectedfrom halogen, —OR⁸, —NO₂, —CN, —CF₃, —OCF₃, —R⁸, thioxo,1,2-methylenedioxy, 1,2-ethylenedioxy, —N(R⁸)₂, —SR⁸, —SOR⁸, —SO₂R⁸,—SO₂N(R⁸)₂, —SO₃R⁸, —C(O)R⁸, —C(O)C(O)R⁸, —C(O)CH₂C(O)R⁸, —C(S)R⁸,—C(S)OR⁸, —C(O)OR⁸, —C(O)C(O)OR⁸, —C(O)C(O)N(R⁸)₂, —OC(O)R⁸,—C(O)N(R⁸)₂, —OC(O)N(R⁸)₂, —C(S)N(R⁸)₂, —(CH₂)₀₋₂NHC(O)R⁸,—N(R⁸)N(R⁸)COR⁸, —N(R⁸)N(R⁸)C(O)OR⁸, —N(R⁸)N(R⁸)CON(R⁸)₂, —N(R⁸)SO₂R⁸,—N(R⁸)SO₂N(R⁸)₂, —N(R⁸)C(O)OR⁸, —N(R⁸)C(O)R⁸, —N(R⁸)C(S)R⁸,—N(R⁸)C(O)N(R⁸)₂, —N(R⁸)C(S)N(R⁸)₂, —N(COR⁸)COR⁸, —N(OR⁸)R⁸,—C(═NH)N(R⁸)₂, —C(O)N(OR⁸)R⁸, —C(═NOR⁸)R⁸, —OP(O)(OR⁸)₂, —P(O)(R⁸)₂,—P(O)(OR⁸)₂, or —P(O)(H)(OR⁸);

each occurrence of R⁸ is independently selected from H— and(C1-C6)-aliphatic-.

In some embodiments, each occurrence of R⁷ is independently selectedfrom —OR⁸, —NO₂, —CN, —CF₃, —OCF₃, —R⁸, thioxo, 1,2-methylenedioxy,1,2-ethylenedioxy, —N(R⁸)₂, —SR⁸, —SOR⁸, —SO₂R⁸, —SO₂N(R⁸)₂, —SO₃R⁸,—C(O)R⁸, —C(O)C(O)R⁸, —C(O)CH₂C(O)R⁸, —C(S)R⁸, —C(S)OR⁸, —C(O)OR⁸,—C(O)C(O)OR⁸, —C(O)C(O)N(R⁸)₂, —OC(O)R⁸, —C(O)N(R⁸)₂, —OC(O)N(R⁸)₂,—C(S)N(R⁸)₂, —(CH₂)₀₋₂NHC(O)R⁸, —N(R⁸)N(R⁸)COR⁸, —N(R⁸)N(R⁸)C(O)OR⁸,—N(R⁸)N(R⁸)CON(R⁸)₂, —N(R⁸)SO₂R⁸, —N(R⁸)SO₂N(R⁸)₂, —N(R⁸)C(O)OR⁸,—N(R⁸)C(O)R⁸, —N(R⁸)C(S)R⁸, —N(R⁸)C(O)N(R⁸)₂, —N(R⁸)C(S)N(R⁸)₂,—N(COR⁸)COR⁸, —N(OR⁸)R⁸, —C(═NH)N(R⁸)₂, —C(O)N(OR⁸)R⁸, —C(═NOR⁸)R⁸,—OP(O)(OR⁸)₂, —P(O)(R⁸)₂, —P(O)(OR⁸)₂, or —P(O)(H)(OR⁸);

each occurrence of R⁸ is independently selected from H— and(C1-C6)-aliphatic-.

According to certain embodiments, the present disclosure provides acompound of formula I, where Y is a C1-aliphatic group optionallysubstituted with one or more R⁴. For example, Y is —CH₂—.

In some embodiments, when Y is a C1-aliphatic group optionallysubstituted with lower alkyl (e.g., C1-C7 alkyl), A is not: (i)unsubstituted phenyl; or

(ii) phenyl substituted with halo (e.g., F— and Cl—) or lower alkyl(e.g., C1-C7 alkyl).

In some embodiments, when Y is a C1-aliphatic group optionallysubstituted with lower alkyl (e.g., C1-C7 alkyl), A is not:

-   (i) unsubstituted phenyl;-   (ii) phenyl substituted with halo (e.g., F— and Cl—) or lower alkyl    (e.g., C1-C7 alkyl); or-   (iii)

In some embodiments, when Y is a C1-aliphatic group optionallysubstituted with lower alkyl (e.g., C1-C7 alkyl), A is not:

-   -   (i) unsubstituted phenyl;    -   (ii) phenyl substituted with halo (e.g., F— and Cl—) or lower        alkyl (e.g., C1-C7 alkyl);    -   (iii)

or

-   -   (iv)

In some embodiments, when Y is a C1-aliphatic group optionallysubstituted with lower alkyl (e.g., C1-C7 alkyl), A is not:

(i) unsubstituted phenyl; (ii) phenyl substituted with halo (e.g., F—and Cl—) or lower alkyl (e.g. C1-C7 alkyl),

(iii)

(iv)

or (v)

andwhen Y is

A is not

According to certain embodiments, the present disclosure provides acompound of formula I, I-A or I-B, as described herein, where Y is aC1-aliphatic or C1-alkylene group optionally substituted with one ormore R⁴. For example, Y is —CH₂—. In some embodiments, when Y is aC1-aliphatic or C1-alkylene group (e.g., —CH₂—) optionally substitutedwith lower alkyl (e.g., C1-C7 alkyl), A is not:

-   -   (i) unsubstituted phenyl;    -   (ii) phenyl substituted with halo (e.g., F— and Cl—) or lower        alkyl (e.g., C1-C7 alkyl).

In some embodiments, the provisos or exceptions outlined above forformula I are embodiments of formula I-A or I-B. In other words, incertain embodiments, the disclosure provides a compound of formula I-Aor I-B, subject to any one or more of the negative provisos set forthabove for formula I.

With respect to formula I, I-A, I-B, or II, it is understood that, ascontext requires, the term “ring” when used for variable A may be usedto refer to “ring system”.

In some embodiments of a compound of formula I, Y is a bond or a(C2-C5)-aliphatic group independently and optionally substituted withone or more R⁴. In some embodiments, Y is a C2-aliphatic groupoptionally substituted with one or more R⁴. In some embodiments, Y is—CH₂—C(R⁴)₂—, such as —CH₂—CH₂—. In other embodiments, Y is—CH₂—C(R⁴)₂—, where each R⁴ is independently selected from halogen. Insome embodiments, Y is —CH₂—C(R⁴)₂—, where both occurrences of R⁴ are—F. In another embodiment, Y is —CH₂—C(R⁴)₂—, where each occurrence ofR⁴ is independently a (C1-C3)-aliphatic group. In yet anotherembodiment, Y is —CH₂—C(R⁴)₂—, where both occurrences of R⁴ are —CH₃.

In some embodiments, the salt is a pharmaceutically acceptable salt of acompound of formula I, such as a sodium salt. In some embodiments, thesalt is a pharmaceutically acceptable salt of a compound of formula I-Aor I-B, such as a sodium salt.

In certain embodiments of compound of formula I, A is a(C5-C10)-aromatic ring having up to 5 heteroatoms independently selectedfrom N, O and S, wherein the aromatic ring is independently andoptionally substituted with one or more R⁷. In some embodiments, A is anoptionally substituted 5- or 6-membered aromatic ring having up to 2heteroatoms selected from N, O and S. In some embodiments, A is anoptionally substituted bi-cyclic aromatic ring having up to 4heteroatoms selected from N, O and S.

In some embodiments, A is a bi-cyclic heteroaryl, such as a bicyclicheteroaryl containing at least N (e.g., containing at least oneheteroatom which is N, such as containing one to three heteroatoms orcontaining up to four heteroatoms, wherein at least one heteroatom isN). In some embodiments, A is an optionally substituted bi-cyclicaromatic ring (e.g., an aromatic ring system) containing at least one N.In some embodiments, A contains at least one N, and when A issubstituted, R⁷ is not an oxo substituent. In some embodiments, A is anoptionally substituted heteroaryl. In some embodiments, A is anoptionally substituted monocylic heteroaryl. In some embodiments, Acontains one to three heteroatoms selected from N, O or S. In someembodiments, A contains at least one N. In some embodiments, A is anoptionally substituted bicyclic heteroaryl, such as a bi-cyclicheteroaryl containing at least one N (e.g., containing at least oneheteroatom which is N, such as containing one to three heteroatoms orcontaining up to four heteroatoms, wherein at least one heteroatom isN).

In some embodiments, A may be an optionally substituted 5- or 6-memberedaromatic group selected from:

wherein A is optionally further substituted with one or more R⁷. In someof these embodiments, Y is a bond or a (C2-C5)-aliphatic groupindependently and optionally substituted with one or more R⁴. In some ofthese embodiments, Y is a C2-aliphatic group independently andoptionally substituted with one or more R⁴. In certain embodiments, A isnot phenyl.

In some embodiments of formula I, A is an optionally substituted 9- or10-membered bicyclic aromatic ring having up to 4 heteroatoms selectedfrom N, O and S. In some embodiments, A is an optionally substitutedbicyclic aromatic ring containing two fused 6-membered aromatic rings,wherein the optionally substituted bicyclic aromatic ring may contain upto 4 nitrogen atoms. In some embodiments, A is an optionally substitutedbicyclic aromatic ring containing one 6-membered aromatic ring fused toone 5-membered aromatic ring, wherein the optionally substitutedbicyclic aromatic ring may contain up to 4 heteroatoms selected from N,O and S. These are examples of ring systems. For example, A may be abicyclic aromatic group (e.g., bicyclic heteroaryl) selected from:

wherein A is optionally further substituted with one or more R⁷. In someembodiments, A is an aromatic group selected from:

wherein A is optionally further substituted with one or more R⁷. In someof these embodiments, Y is a bond or a (C2-C5)-aliphatic groupindependently and optionally substituted with one or more independentlyselected R⁴. In some of these embodiments, Y is a C2-aliphatic groupindependently and optionally substituted with one or more independentlyselected R⁴.

In certain embodiments, A is a bicyclic aromatic ring (e.g., bicyclicheteroaryl) selected from:

wherein A is optionally further substituted with one or more R⁷. Incertain embodiment, A is a bicyclic aromatic group (e.g., a ring system)(e.g., bicyclic heteroaryl) selected from:

wherein A is optionally further substituted with one or more R⁷.

In certain embodiments, A is selected from:

wherein A is optionally further substituted with one or more R⁷.

In certain embodiments, A is selected from:

wherein A is optionally further substituted with one or more R⁷.

In certain embodiments, A is selected from:

wherein A is optionally further substituted with one or more R⁷.

In some embodiments, A is selected from:

optionally further substituted with one or more R⁷.

In some embodiments, A is

optionally further substituted with one or more R⁷.

In certain embodiments, A is

optionally further substituted with one or more R⁷. In certainembodiments, A is

optionally further substituted with one or more R⁷, wherein R⁸ is notmethyl.

In another embodiment, A is

optionally substituted with one or more R⁷. In some of the aboveembodiments of A, each occurrence of R⁷ is independently selected fromhalogen, —CF₃, —OCF₃, —C1-C4 aliphatic (e.g., —C1-C4 alkyl), and—O(C1-C4 aliphatic) (e.g., —O(C1-C4 alkyl)).

In certain embodiments, the present disclosure provides compounds offormula I, where X is —H, —C(O)R⁵, or —C(O)OR⁵. In some embodiments, Xis —H. In other embodiments, X is a prodrug moiety that is cleaved underphysiological conditions to provide a compound wherein X is H. In someembodiments, X is —P(O)(OR⁵)₂. In some embodiments, X is a prodrugmoiety that is cleaved under physiological conditions to provide acompound wherein X is —P(O)(OR⁵)₂, such as PO₃H₂, or a salt thereof.

In some embodiments, the present disclosure also provides compounds offormula I, where R¹ is —H, bromine, iodine, methyl, ethyl or —CF₃. Insome embodiments, R¹ is —H.

According to certain embodiments, the present disclosure provides acompound of formula I, where Z is ═O or ═S. In some embodiments, Z is═O.

In some embodiments, the compound of the present disclosure has a W thatis ═O or ═S. In some embodiments, W is ═O.

In some embodiments, the present disclosure provides a compound offormula I, where R² and R³ are each independently —OR⁵. In someembodiments, R² is —OH. In another embodiment, R³ is —OH.

The disclosure also includes various combinations of A, X, Y, Z, W, R¹,R² and R³ as described above. These combinations can in turn be combinedwith any or all of the values of the other variables described above.For example, in some embodiments, Y is a C1- or C2-aliphatic groupoptionally substituted with one or more R⁴ and X is —H, —C(O)R⁵, or—C(O)OR⁵ ₂. In another embodiment, Y is a C1- or C2-aliphatic groupoptionally substituted with one or more R⁴; X is —H, —C(O)R⁵, or—C(O)OR⁵; and Z is ═O. In another embodiment, Y is a C1- or C2-aliphaticgroup optionally substituted with one or more R⁴; X is —H, —C(O)R⁵, or—C(O)OR⁵ ₂; Z is ═O; and W is ═O. In yet another embodiment, Y is a C1-or C2-aliphatic group optionally substituted with one or more R⁴; X is—H, —C(O)R⁵, or —C(O)OR⁵; Z is ═O; W is ═O; and R¹ is selected from —H,bromine, iodine, methyl, ethyl, and —CF₃, for example, R¹ is —H. In afurther embodiment, Y is a C1- or C2-aliphatic group optionallysubstituted with one or more R⁴; X is —H, —C(O)R⁵, or —C(O)OR⁵; Z is ═O;W is ═O; and R¹ is selected from —H, bromine, iodine, methyl, ethyl, and—CF₃; and A is selected from the following groups:

wherein A is optionally further substituted with one or more R⁷, forexample, A is optionally substituted

In a further embodiment, Y is a C1- or C2-aliphatic group optionallysubstituted with one or more R⁴; X is —H, —C(O)R⁵, or —C(O)OR⁵—; Z is═O; W is ═O; and R¹ is selected from —H, bromine, iodine, methyl, ethyl,and —CF₃; A is selected from the following group:

wherein A is optionally further substituted with one or more R⁷; and R²and R³ are each independently —OR⁵, for example, R² and R³ are eachindependently —OH. In some of the above embodiments, A is

optionally further substituted with one or more R⁷. In some of the aboveembodiments, A is

optionally further substituted with one or more R⁷, wherein R⁸ is notmethyl. In another embodiment, Y is a C1- or C2-aliphatic groupoptionally substituted with one or more R⁴; X is —H, —C(O)R⁵, or—C(O)OR⁵; Z is ═O; W is ═O; and R¹ is selected from —H, bromine, iodine,methyl, ethyl, and —CF₃; and A is selected from the following groups:

wherein A is optionally further substituted with one or more R⁷, forexample, A is optionally substituted

In a further embodiment, Y is a C1- or C2-aliphatic group optionallysubstituted with one or more R⁴; X is —H, —C(O)R⁵, or —C(O)OR⁵; Z is ═O;W is ═O; and R¹ is selected from —H, bromine, iodine, methyl, ethyl, and—CF₃; A is selected from the following groups:

wherein A is optionally further substituted with one or more R⁷;and R² and R³ are each independently —OR⁵, for example, R² and R³ areeach independently —OH. In some of the above embodiments, when Y is aC1-aliphatic group optionally substituted with lower alkyl (e.g., C1-C7alkyl), A is not:

-   -   (i) unsubstituted phenyl;    -   (ii) phenyl substituted with halo (e.g., F— and Cl—) or lower        alkyl (e.g., C1-C7 alkyl);    -   (iii)

or

-   -   (iv)

In some embodiments, when Y is a C1-aliphatic group optionallysubstituted with lower alkyl (e.g., C1-C7 alkyl), A is not:

(i) unsubstituted phenyl; (ii) phenyl substituted with halo (e.g., F—and Cl—) or lower alkyl (e.g., C1-C7 alkyl),

(iii)

(iv)

(v)

andwhen Y is

A is not

In some of the above embodiments, Y is a C2-aliphatic groupindependently and optionally substituted with one or more independentlyselected R⁴. In some of the above embodiments, each occurrence of R⁷ isindependently selected from halogen, —CF₃, —OCF₃, —C1-C4 aliphatic(e.g., —C1-C4 alkyl), and —O(C1-C4 aliphatic) (e.g., —O(C1-C4 alkyl)).

The present disclosure also provides a compound of formula I-A:

-   or a salt thereof, wherein:-   A is a 3- to 10-membered aromatic or non-aromatic ring having up to    5 heteroatoms independently selected from N, O, S, SO, or SO₂,    wherein the aromatic or non-aromatic ring is independently and    optionally substituted with one or more R⁷;-   X is selected from:    -   —H, —C(O)R⁵, —C(O)OR⁵, and —P(O)(OR⁵)₂, preferably X is —H,        —C(O)R⁵, or —C(O)OR⁵;-   Y is a bond or a (C1-C5)-aliphatic group independently and    optionally substituted with one or more R⁴;-   Z and W are each independently selected from ═O, ═S, ═N(R⁵), and    ═NOR⁵;-   R¹ is selected from:    -   —H, halogen, —OR⁵, —CN, —CF₃, —OCF₃ and a (C1-C6)-aliphatic        group optionally substituted with one or more R⁷;-   R² and R³ are each independently selected from —OR⁵, —SR⁵, —NR⁵R⁶,    —OC(O)R⁵, —OC(O)NR⁵R⁶, and —OC(O)OR⁵; preferably, R² and R³ are each    independently selected from —OR⁵, —SR⁵, —NR⁵R⁶ and —OC(O)R⁵;-   each occurrence of R⁴ is independently selected from:    -   halogen, —OR⁵, —NO₂, —CN, —CF₃, —OCF₃, —R⁵, 1,2-methylenedioxy,        1,2-ethylenedioxy, —N(R⁵)₂, —SR⁵, —SOR⁵, —SO₂R⁵, —SO₂N(R⁵)₂,        —SO₃R⁵, —C(O)R⁵, —C(O)C(O)R⁵, —C(O)CH₂C(O)R⁵, —C(S)R⁵, —C(S)OR⁵,        —C(O)OR⁵, —C(O)C(O)OR⁵, —C(O)C(O)N(R⁵)₂, —OC(O)R⁵, —C(O)N(R⁵)₂,        —OC(O)N(R⁵)₂, —C(S)N(R⁵)₂, —(CH₂)₀₋₂NHC(O)R⁵, —N(R⁵)N(R⁵)COR⁵,        —N(R⁵)N(R⁵)C(O)OR⁵, —N(R⁵)N(R⁵)CON(R⁵)₂, —N(R⁵)SO₂R⁵,        —N(R⁵)SO₂N(R⁵)₂, —N(R⁵)C(O)OR⁵, —N(R⁵)C(O)R⁵, —N(R⁵)C(S)R⁵,        —N(R⁵)C(O)N(R⁵)₂, —N(R⁵)C(S)N(R⁵)₂, —N(COR⁵)COR⁵, —N(OR⁵)R⁵,        —C(═NH)N(R⁵)₂, —C(O)N(OR⁵)R⁵, —C(═NOR⁵)R⁵, —OP(O)(OR⁵)₂,        —P(O)(R⁵)₂, —P(O)(OR⁵)₂, or —P(O)(H)(OR⁵);-   each occurrence of R⁵ is independently selected from:    -   H—,    -   (C1-C12)-aliphatic-,    -   (C3-C10)-cycloalkyl- or -cycloalkenyl-,    -   [(C3-C10)-cycloalkyl or -cycloalkenyl]-(C1-C12)-aliphatic-,    -   (C6-C10)-aryl-,    -   (C6-C10)-aryl-(C1-C12)aliphatic-,    -   (C3-C10)-heterocyclyl-,    -   (C6-C10)-heterocyclyl-(C1-C12)aliphatic-,    -   (C5-C10)-heteroaryl-, and    -   (C5-C10)-heteroaryl-(C1-C12)-aliphatic-;    -   wherein two R⁵ groups bound to the same atom optionally form a        3- to 10-membered aromatic or non-aromatic ring having up to 3        heteroatoms independently selected from N, O, S, SO, or SO₂,        wherein said ring is optionally fused to a (C6-C10)aryl,        (C5-C10)heteroaryl, (C3-C10)cycloalkyl, or a        (C3-C10)heterocyclyl; and    -   wherein each R⁵ group is independently and optionally        substituted with one or more R⁷;-   R⁶ is selected from:    -   —R⁵, —C(O)R⁵, —C(O)OR⁵, —C(O)N(R⁵)₂ and —S(O)₂R⁵;-   each occurrence of R⁷ is independently selected from:    -   halogen, —OR⁸, —NO₂, —CN, —CF₃, —OCF₃, —R⁸, oxo, thioxo,        1,2-methylenedioxy, 1,2-ethylenedioxy, —N(R⁸)₂, —SR⁸, —SOR⁸,        —SO₂R⁸, —SO₂N(R⁸)₂, —SO₃R⁸, —C(O)R⁸, —C(O)C(O)R⁸,        —C(O)CH₂C(O)R⁸, —C(S)R⁸, —C(S)OR⁸, —C(O)OR⁸, —C(O)C(O)OR⁸,        —C(O)C(O)N(R⁸)₂, —OC(O)R⁸, —C(O)N(R⁸)₂, —OC(O)N(R⁸)₂,        —C(S)N(R⁸)₂, —(CH₂)₀₋₂NHC(O)R⁸, —N(R⁸)N(R⁸)COR⁸,        —N(R⁸)N(R⁸)C(O)OR⁸, —N(R⁸)N(R⁸)CON(R⁸)₂, —N(R⁸)SO₂R⁸,        —N(R⁸)SO₂N(R⁸)₂, —N(R⁸)C(O)OR⁸, —N(R⁸)C(O)R⁸, —N(R⁸)C(S)R⁸,        —N(R⁸)C(O)N(R⁸)₂, —N(R⁸)C(S)N(R⁸)₂, —N(COR⁸)COR⁸, —N(OR⁸)R⁸,        —C(═NH)N(R⁸)₂, —C(O)N(OR⁸)R⁸, —C(═NOR⁸)R⁸, —OP(O)(OR⁸)₂,        —P(O)(R⁸)₂, —P(O)(OR⁸)₂, or —P(O)(H)(OR⁸);-   each occurrence of R⁸ is independently selected from:    -   H— and (C1-C6)-aliphatic-;-   provided that A is not

In certain embodiments, the present disclosure provides compounds offormula I-A where the definition of one or more of variables A, X, Y, Z,W, and R¹-R⁸ is as defined by embodiments described above for Formula I,including any of the combinations thereof, as well as the provisosprovided for Formula I.

According to certain embodiments, the present disclosure provides acompound of formula I-A, where Y is a C1-aliphatic group optionallysubstituted with one or more R⁴. For example, Y is —CH₂—. In someembodiments, when Y is a C1-aliphatic group optionally substituted withlower alkyl (e.g., C1-C7 alkyl), A is not:

-   -   (i) unsubstituted phenyl; or    -   (ii) phenyl substituted with halo (e.g., F— and Cl—) or lower        alkyl (e.g., C1-C7 alkyl).

The present disclosure also provides a compound of formula I-B:

-   or a salt thereof, wherein:-   A is selected from:    -   a phenyl group;    -   a naphthalene group;    -   a 5- to 10-membered heteroaryl group having up to 5 heteroatoms        independently selected from N, O, and S; and    -   a 3- to 10-membered non-aromatic ring having up to 5 heteroatoms        independently selected from N, O, and S;    -   wherein A is optionally further substituted with 1, 2, or 3        substituents independently selected from the group consisting of        halogen, hydroxyl, (C1-C6)-alkyl, and (C1-C6)-alkoxyl;-   Y is a (C1-C6)-alkylene optionally substituted with halogen; and-   R¹ is —H, halogen, or a (C1-C6)-aliphatic group optionally    substituted with one or more halogen.

In certain embodiments, A is a 5- to 10-membered heteroaryl group havingup to 3 heteroatoms independently selected from N, O, and S. In certainother embodiments, A is not

In certain other embodiments, R¹ is hydrogen or methyl. In certain otherembodiments, the present disclosure provides compounds of formula I-B,wherein Y is (C1-C5)-alkylene optionally substituted with halogen.

According to certain embodiments, the present disclosure provides acompound of formula I-B, where Y is a C1-alkylene group (for example, Yis —CH₂—), which is optionally substituted with one or more R⁴. In someembodiments, when Y is a C1-alkylene group optionally substituted withlower alkyl (e.g., C1-C7 alkyl), A is not:

-   -   (i) unsubstituted phenyl; or    -   (ii) phenyl substituted with halo (e.g., F— and Cl—) or lower        alkyl (e.g., C1-C7 alkyl).

In certain embodiments, when Y is a Cl-aliphatic group optionallysubstituted with lower alkyl, A is not:

-   (i) unsubstituted phenyl;-   (ii) phenyl substituted with halo or lower alkyl; or-   (iii)

In certain embodiments, when Y is a Cl-aliphatic group optionallysubstituted with lower alkyl, A is not:

-   (i) unsubstituted phenyl;-   (ii) phenyl substituted with halo or lower alkyl;-   (iii)

(iv)

or (v)

and

-   when Y is

A is not

In some embodiments of formula I-A and I-B, A is an optionallysubstituted 9- or 10-membered bicyclic aromatic ring having up to 4heteroatoms selected from N, O and S. In some embodiments, A is anoptionally substituted bicyclic aromatic ring containing two fused6-membered aromatic rings, wherein the optionally substituted bicyclicaromatic ring may contain up to 4 nitrogen atoms. In some embodiments, Ais an optionally substituted bicyclic aromatic ring containing one6-membered aromatic ring fused to one 5-membered aromatic ring, whereinthe optionally substituted bicyclic aromatic ring may contain up to 4heteroatoms selected from N, O and S. In certain embodiments, A is

optionally further substituted with one or more R⁷. In certainembodiments, A is

optionally further substituted with one or more R⁷. In certain otherembodiments, A is

In certain other embodiments, A is not

In some embodiments, A contains at least one N, and when A issubstituted, R⁷ is not an oxo substituent. In some embodiments, A is anoptionally substituted heteroaryl. In some embodiments, A is anoptionally substituted monocylic heteroaryl. In some embodiments, Acontains one to three heteroatoms selected from N, O or S. In someembodiments, A contains at least one N. In some embodiments, A is anoptionally substituted bicyclic heteroaryl. In some embodiments, Acontains at least one N.

The present disclosure also provides a compound of formula II:

-   or a prodrug or salt thereof, wherein:-   A is selected from:    -   a phenyl group;    -   a naphthalene group;    -   a 5- to 10-membered heteroaryl group having up to 5 heteroatoms        independently selected from N, O and S; and    -   a 3- to 10-membered non-aromatic ring having up to 5 heteroatoms        independently selected from N, O, S, SO, or SO₂;    -   wherein A is optionally further substituted with one or more R⁷;-   X is selected from:    -   —H, —C(O)R⁵, —C(O)OR⁵, and —P(O)(OR⁵)₂, preferably X is —H,        —C(O)R⁵, or —C(O)OR⁵;-   Y¹ is a (C1-C5)-aliphatic group substituted with at least one oxo    and further independently and optionally substituted with one or    more R⁴;-   Z and W are each independently selected from ═O, ═S, ═N(R⁵), and    ═NOR⁵;-   R¹ is selected from:    -   —H, halogen, —OR⁵, —CN, —CF₃, —OCF₃ and a (C1-C6)-aliphatic-        group optionally substituted with one or more R⁴;-   R² and R³ are each independently selected from —OR⁵, —SR⁵, —NR⁵R⁶,    —OC(O)R⁵, —OC(O)NR⁵R⁶, and —OC(O)OR⁵; preferably, R² and R³ are each    independently selected from —OR⁵, —SR⁵, —NR⁵R⁶ and —OC(O)R⁵;-   each occurrence of R⁴ is independently selected from:    -   halogen, —OR⁵, —NO₂, —CN, —CF₃, —OCF₃, —R⁵, oxo, thioxo,        1,2-methylenedioxy, 1,2-ethylenedioxy, —N(R⁵)₂, —SR⁵, —SOR⁵,        —SO₂R⁵, —SO₂N(R⁵)₂, —SO₃R⁵, —C(O)R⁵, —C(O)C(O)R⁵,        —C(O)CH₂C(O)R⁵, —C(S)R⁵, —C(S)OR⁵, —C(O)OR⁵, —C(O)C(O)OR⁵,        —C(O)C(O)N(R⁵)₂, —OC(O)R⁵, —C(O)N(R⁵)₂, —OC(O)N(R⁵)₂,        —C(S)N(R⁵)₂, —(CH₂)₀₋₂NHC(O)R⁵, —N(R⁵)N(R⁵)COR⁵,        —N(R⁵)N(R⁵)C(O)OR⁵, —N(R⁵)N(R⁵)CON(R⁵)₂, —N(R⁵)SO₂R⁵,        —N(R⁵)SO₂N(R⁵)₂, —N(R⁵)C(O)OR⁵, —N(R⁵)C(O)R⁵, —N(R⁵)C(S)R⁵,        —N(R⁵)C(O)N(R⁵)₂, —N(R⁵)C(S)N(R⁵)₂, —N(COR⁵)COR⁵, —N(OR⁵)R⁵,        —C(═NH)N(R⁵)₂, —C(O)N(OR⁵)R⁵, —C(═NOR⁵)R⁵, —OP(O)(OR⁵)₂,        —P(O)(R⁵)₂, —P(O)(OR⁵)₂, or —P(O)(H)(OR⁵);-   each occurrence of R⁵ is independently selected from:    -   H—,    -   (C1-C12)-aliphatic-,    -   (C3-C10)-cycloalkyl- or -cycloalkenyl-,    -   [(C3-C10)-cycloalkyl or -cycloalkenyl]-(C1-C12)-aliphatic-,    -   (C6-C10)-aryl-,    -   (C6-C10)-aryl-(C1-C12)aliphatic-,    -   (C3-C10)-heterocyclyl-,    -   (C6-C10)-heterocyclyl-(C1-C12)aliphatic-,    -   (C5-C10)-heteroaryl-, and    -   (C5-C10)-heteroaryl-(C1-C12)-aliphatic-;    -   wherein two R⁵ groups bound to the same atom optionally form a        3- to 10-membered aromatic or non-aromatic ring having up to 3        heteroatoms independently selected from N, O, S, SO, or SO₂,        wherein said ring is optionally fused to a (C6-C10)aryl,        (C5-C10)heteroaryl, (C3-C10)cycloalkyl, or a        (C3-C10)heterocyclyl; and    -   wherein each R⁵ group is independently and optionally        substituted with one or more R⁷;-   R⁶ is selected from:    -   —R⁵, —C(O)R⁵, —C(O)OR⁵, —C(O)N(R⁵)₂ and —S(O)₂R⁵;-   each occurrence of R⁷ is independently selected from:    -   halogen, —OR⁸, —NO₂, —CN, —CF₃, —OCF₃, —R⁸, oxo, thioxo,        1,2-methylenedioxy, 1,2-ethylenedioxy, —N(R⁸)₂, —SR⁸, —SOR⁸,        —SO₂R⁸, —SO₂N(R⁸)₂, —SO₃R⁸, —C(O)R⁸, —C(O)C(O)R⁸,        —C(O)CH₂C(O)R⁸, —C(S)R⁸, —C(S)OR⁸, —C(O)OR⁸, —C(O)C(O)OR⁸,        —C(O)C(O)N(R⁸)₂, —OC(O)R⁸, —C(O)N(R⁸)₂, —OC(O)N(R⁸)₂,        —C(S)N(R⁸)₂, —(CH₂)₀₋₂NHC(O)R⁸, —N(R⁸)N(R⁸)COR⁸,        —N(R⁸)N(R⁸)C(O)OR⁸, —N(R⁸)N(R⁸)CON(R⁸)₂, —N(R⁸)SO₂R⁸,        —N(R⁸)SO₂N(R⁸)₂, —N(R⁸)C(O)OR⁸, —N(R⁸)C(O)R⁸, —N(R⁸)C(S)R⁸,        —N(R⁸)C(O)N(R⁸)₂, —N(R⁸)C(S)N(R⁸)₂, —N(COR⁸)COR⁸, —N(OR⁸)R⁸,        —C(═NH)N(R⁸)₂, —C(O)N(OR⁸)R⁸, —C(═NOR⁸)R⁸, —OP(O)(OR⁸)₂,        —P(O)(R⁸)₂, —P(O)(OR⁸)₂, or —P(O)(H)(OR⁸);-   each occurrence of R⁸ is independently selected from:    -   H— and (C1-C6)-aliphatic-.

In some embodiments of compound of formula II, each occurrence of R⁴ isindependently selected from:

-   -   halogen, —OR⁵, —NO₂, —CN, —CF₃, —OCF₃, —R⁵, oxo, thioxo,        —N(R⁵)₂, —SR⁵, —SOR⁵, —SO₂R⁵, —SO₂N(R⁵)₂, —SO₃R⁵, —C(O)R⁵,        —C(O)C(O)R⁵, —C(O)CH₂C(O)R⁵, —C(S)R⁵, —C(S)OR⁵, —C(O)OR⁵,        —C(O)C(O)OR⁵, —C(O)C(O)N(R⁵)₂, —OC(O)R⁵, —C(O)N(R⁵)₂,        —OC(O)N(R⁵)₂, —C(S)N(R⁵)₂, —(CH₂)₀₋₂NHC(O)R⁵, —N(R⁵)N(R⁵)COR⁵,        —N(R⁵)N(R⁵)C(O)OR⁵, —N(R⁵)N(R⁵)CON(R⁵)₂, —N(R⁵)SO₂R⁵,        —N(R⁵)SO₂N(R⁵)₂, —N(R⁵)C(O)OR⁵, —N(R⁵)C(O)R⁵, —N(R⁵)C(S)R⁵,        —N(R⁵)C(O)N(R⁵)₂, —N(R⁵)C(S)N(R⁵)₂, —N(COR⁵)COR⁵, —N(OR⁵)R⁵,        —C(═NH)N(R⁵)₂, —C(O)N(OR⁵)R⁵, —C(═NOR⁵)R⁵, —OP(O)(OR⁵)₂,        —P(O)(R⁵)₂, —P(O)(OR⁵)₂, or —P(O)(H)(OR⁵).

In some embodiments, R⁷ is independently selected from halogen, —OR⁸,—NO₂, —CN, —CF₃, —OCF₃, —R⁸, thioxo, 1,2-methylenedioxy,1,2-ethylenedioxy, —N(R⁸)₂, —SR⁸, —SOR⁸, —SO₂R⁸, —SO₂N(R⁸)₂, —SO₃R⁸,—C(O)R⁸, —C(O)C(O)R⁸, —C(O)CH₂C(O)R⁸, —C(S)R⁸, —C(S)OR⁸, —C(O)OR⁸,—C(O)C(O)OR⁸, —C(O)C(O)N(R⁸)₂, —OC(O)R⁸, —C(O)N(R⁸)₂, —OC(O)N(R⁸)₂,—C(S)N(R⁸)₂, —(CH₂)₀₋₂NHC(O)R⁸, —N(R⁸)N(R⁸)COR⁸, —N(R⁸)N(R⁸)C(O)OR⁸,—N(R⁸)N(R⁸)CON(R⁸)₂, —N(R⁸)SO₂R⁸, —N(R⁸)SO₂N(R⁸)₂, —N(R⁸)C(O)OR⁸,—N(R⁸)C(O)R⁸, —N(R⁸)C(S)R⁸, —N(R⁸)C(O)N(R⁸)₂, —N(R⁸)C(S)N(R⁸)₂,—N(COR⁸)COR⁸, —N(OR⁸)R⁸, —C(═NH)N(R⁸)₂, —C(O)N(OR⁸)R⁸, —C(═NOR⁸)R⁸,—OP(O)(OR⁸)₂, —P(O)(R⁸)₂, —P(O)(OR⁸)₂, or —P(O)(H)(OR⁸);

-   each occurrence of R⁸ is independently selected from H— and    (C1-C6)-aliphatic-.

In some embodiments, R⁷ is independently selected from —OR⁸, —NO₂, —CN,—CF₃, —OCF₃, —R⁸, thioxo, 1,2-methylenedioxy, 1,2-ethylenedioxy,—N(R⁸)₂, —SR⁸, —SOR⁸, —SO₂R⁸, —SO₂N(R⁸)₂, —SO₃R⁸, —C(O)R⁸, —C(O)C(O)R⁸,—C(O)CH₂C(O)R⁸, —C(S)R⁸, —C(S)OR⁸, —C(O)OR⁸, —C(O)C(O)OR⁸,—C(O)C(O)N(R⁸)₂, —OC(O)R⁸, —C(O)N(R⁸)₂, —OC(O)N(R⁸)₂, —C(S)N(R⁸)₂,—(CH₂)₀₋₂NHC(O)R⁸, —N(R⁸)N(R⁸)COR⁸, —N(R⁸)N(R⁸)C(O)OR⁸,—N(R⁸)N(R⁸)CON(R⁸)₂, —N(R⁸)SO₂R⁸, —N(R⁸)SO₂N(R⁸)₂, —N(R⁸)C(O)OR⁸,—N(R⁸)C(O)R⁸, —N(R⁸)C(S)R⁸, —N(R⁸)C(O)N(R⁸)₂, —N(R⁸)C(S)N(R⁸)₂,—N(COR⁸)COR⁸, —N(OR⁸)R⁸, —C(═NH)N(R⁸)₂, —C(O)N(OR⁸)R⁸, —C(═NOR⁸)R⁸,—OP(O)(OR⁸)₂, —P(O)(R⁸)₂, —P(O)(OR⁸)₂, or —P(O)(H)(OR⁸)

-   each occurrence of R⁸ is independently selected from H— and    (C1-C6)-aliphatic-.

According to certain embodiments, the present disclosure also provides acompound of formula II, where Y¹ is a C1-aliphatic group substitutedwith oxo. In some embodiments, Y¹ is a C2-aliphatic group substitutedwith at least one oxo and optionally further substituted with one ormore R⁴. In another embodiment, Y¹ is —C(O)—C(R⁴)₂— or —C(R⁴)₂—C(O)—,for example, —C(O)—CH₂— or —CH₂—C(O)—. In a further embodiment, Y¹ is—C(O)—C(R⁴)₂— or —C(R⁴)₂—C(O)—, where each R⁴ is independently selectedfrom halogen. For example, Y¹ is —C(O)—C(R⁴)₂— or —C(R⁴)₂—C(O)—, whereboth occurrences of R⁴ in are —F. In yet another embodiment, Y¹ is—C(O)—C(R⁴)₂— or —C(R⁴)₂—C(O)—, where each R⁴ is independently a(C1-C3)-aliphatic group. For example, Y¹ is —C(O)—C(R⁴)₂— or—C(R⁴)₂—C(O)—, where both occurrences of R⁴ are —CH₃. In some of theabove embodiments, when Y¹ is a —CH₂—C(O)—, wherein the carbonyl groupof Y¹ is connected directly to A, A is not:

-   -   (i) unsubstituted phenyl; or    -   (ii) phenyl substituted with halo (e.g., F— and Cl—) or lower        alkyl (e.g., C1-C7 alkyl);

In some embodiments, the salt is a pharmaceutically acceptable salt of acompound of formula II, such as a sodium salt.

In certain embodiments of compound of formula II, A is a phenyl group; anaphthalene group; or a 5- to 10-membered heteroaryl group having up to5 heteroatoms independently selected from N, O and S, wherein A isoptionally substituted with one or more R⁷. For example, A is selectedfrom the following groups:

where A is optionally substituted with one or more R⁷. In certainembodiments, A is not a phenyl group.

In certain embodiments of compound of formula II, A is a(C5-C10)-aromatic ring having up to 5 heteroatoms independently selectedfrom N, O and S, wherein the aromatic ring is independently andoptionally substituted with one or more R⁴. In some embodiments, A is anoptionally substituted 5- or 6-membered aromatic ring having up to 2heteroatoms selected from N, O and S. For example, A is an aromaticgroup selected from:

wherein A is optionally further substituted with one or more R⁷. In someof these embodiments, Y¹ is a C1-aliphatic or a C3-C5-aliphatic groupsubstituted with at least one oxo and further independently andoptionally substituted with one or more independently selected R⁴;

In some embodiments of formula II, A is an optionally substituted 9- or10-membered bicyclic aromatic ring having up to 4 heteroatoms selectedfrom N, O and S. In some embodiments, A is an optionally substitutedbicyclic aromatic ring containing two fused 6-membered aromatic rings,wherein the optionally substituted bicyclic aromatic ring may contain upto 4 nitrogen atoms. In some embodiments, A is an optionally substitutedbicyclic aromatic ring containing one 6-membered aromatic ring fused toone 5-membered aromatic ring, wherein the optionally substitutedbicyclic aromatic ring may contain up to 4 heteroatoms selected from N,O and S. For example, A may be a bicyclic aromatic group selected from:

wherein A is optionally further substituted with one or more R⁷.

In some embodiments of compound of formula II, A is selected from thefollowing groups:

where A is optionally substituted with one or more R⁷.

In such embodiments, A is one of the following groups:

where A is optionally further substituted with one or more R⁷. In someof these embodiments, Y is a C1-aliphatic or a C3-C5-aliphatic groupsubstituted with at least one oxo and further independently andoptionally substituted with one or more independently selected R⁴;

In some embodiments, A is a bicyclic aromatic group selected from:

wherein A is optionally further substituted with one or more R⁷.

In some embodiments, A is selected from:

where A is optionally further substituted with one or more R⁷.

In some embodiments, A is selected from:

where A is optionally further substituted with one or more R⁷.

In some embodiments, A is

where A is optionally further substituted with one or more R⁷.

In some embodiments, A is

where A is optionally further substituted with one or more R⁷.

In some embodiments, A is

optionally further substituted with one or more R⁷. In some embodiments,A is

optionally further substituted with one or more R⁷, wherein R⁸ is notmethyl.

In a further embodiment, A is

optionally substituted with one or more R⁷. In some of the aboveembodiments of A, each occurrence of R⁴ is independently selected fromhalogen, —CF₃, —OCF₃, —C1-C4 aliphatic (e.g., —C1-C4 alkyl), and—O(C1-C4 aliphatic) (e.g., —O(C1-C4 alkyl)).

In some embodiments, Y¹ is a C2-aliphatic group substituted with atleast one oxo and optionally further substituted with one or more R⁴,and A is selected from:

a phenyl group;

a naphthalene group; and

a 6-membered monocyclic or a 9- to 10-membered bicyclic heteroaryl grouphaving up to 5 heteroatoms independently selected from N, O and S,wherein the bicyclic heteroaryl group has a 6-membered aryl orheteroaryl ring that is directly connected to Y¹;

wherein A is optionally further substituted with one or more R⁷. In somesuch embodiments, Y¹ is a C2-aliphatic group substituted with one oxo,and A is selected from:

wherein A is optionally further substituted with one or more R⁷. In someof the above embodiments, when Y¹ is a —CH₂—C(O)—, wherein the carbonylgroup of Y¹ is connected directly to A, A is not:

(i) unsubstituted phenyl; or

(ii) phenyl substituted with halo (e.g., F— and Cl—) or lower alkyl(e.g., C1-C7 alkyl)

According to certain embodiments, the present disclosure provides acompound of formula II, where X is —H, —C(O)R⁵, or —C(O)OR⁵. In someembodiments, X is —H. In other embodiments, X is a prodrug moiety thatis cleaved under physiological conditions to provide a compound whereinX is H. In some embodiments, X is —P(O)(OR⁵)₂. In some embodiments, X isa prodrug moiety that is cleaved under physiological conditions toprovide a compound wherein X is —P(O)(OR⁵)₂, such as PO₃H₂, or a saltthereof.

In some embodiments of the compound of formula II, R¹ is —H, bromine,iodine, methyl, ethyl or —CF₃. In some embodiments, R¹ is —H.

According to certain embodiments, the present disclosure also provides acompound of formula II, where Z is ═O or ═S. In some embodiments, Z is═O.

In some embodiments of the compound of formula II, W is ═O or ═S. Insome embodiments, W is ═O.

In some embodiments of compound of formula II, R² and R³ are eachindependently —OR⁵. In some embodiments, R² is —OH. In anotherembodiment, R³ is —OH.

The disclosure also includes various combinations of A, X, Y¹, Z, W, R¹,R² and R³ as described above. These combinations can in turn be combinedwith any or all of the values of the other variables described above.For example, in some embodiments, Y¹ is a C1-aliphatic group substitutedwith an oxo or a C2-aliphatic group substituted with at least one oxoand optionally further substituted with one or more R⁴ and X is —H,—C(O)R⁵, or —C(O)OR⁵. In another embodiment, Y¹ is a C1-aliphatic groupsubstituted with an oxo or a C2-aliphatic group substituted with atleast one oxo and optionally further substituted with one or more R⁴; Xis —H, —C(O)R⁵, or —C(O)OR⁵; and Z is ═O. In another embodiment, Y¹ is aC1-aliphatic group substituted with an oxo or a C2-aliphatic groupsubstituted with at least one oxo and optionally further substitutedwith one or more R⁴; X is —H, —C(O)R⁵, or —C(O)OR⁵; Z is ═O; and W is═O. In yet another embodiment, Y¹ is a C1-aliphatic group substitutedwith an oxo or a C2-aliphatic group substituted with at least one oxoand optionally further substituted with one or more R⁴; X is —H,—C(O)R⁵, or —C(O)OR⁵; Z is ═O; W is ═O; and R¹ is selected from —H,bromine, iodine, methyl, ethyl, and —CF₃, for example, R¹ is —H. In afurther embodiment, Y¹ is a C1-aliphatic group substituted with an oxoor a C2-aliphatic group substituted with at least one oxo and optionallyfurther substituted with one or more R⁴; X is —H, —C(O)R⁵, or —C(O)OR⁵;Z is ═O; W is ═O; and R¹ is selected from —H, bromine, iodine, methyl,ethyl, and —CF₃; and A is selected from the following groups:

wherein A is optionally further substituted with one or more R⁷, forexample, A is optionally further substituted

In yet a further embodiment, Y¹ is a C1-aliphatic group substituted withan oxo or a C2-aliphatic group substituted with at least one oxo andoptionally further substituted with one or more R⁴; X is —H, —C(O)R⁵, or—C(O)OR⁵; Z is ═O; W is ═O; and R¹ is selected from —H, bromine, iodine,methyl, ethyl, and —CF₃; A is selected from the following group:

wherein A is optionally further substituted with one or more R⁷;and R² and R³ are each independently —OR⁵, for example R² and R³ areeach independently —OH. In some of the above embodiments, A is

optionally further substituted with one or more R⁴. In some of the aboveembodiments, A is

optionally further substituted with one or more R⁴, wherein R⁸ is notmethyl. In a further embodiment, Y¹ is a C1-aliphatic group substitutedwith an oxo or a C2-aliphatic group substituted with at least one oxoand optionally further substituted with one or more R⁴; X is —H,—C(O)R⁵, or —C(O)OR⁵; Z is ═O; W is ═O; and R¹ is selected from —H,bromine, iodine, methyl, ethyl, and —CF₃; and A is selected from thefollowing groups:

wherein A is optionally further substituted with one or more R⁷, forexample, A is optionally further substituted

In yet a further embodiment, Y¹ is a C1-aliphatic group substituted withan oxo or a C2-aliphatic group substituted with at least one oxo andoptionally further substituted with one or more R⁴; X is —H, —C(O)R⁵, or—C(O)OR⁵; Z is ═O; W is ═O; and R¹ is selected from —H, bromine, iodine,methyl, ethyl, and —CF₃; A is selected from the following group:

wherein A is optionally further substituted with one or more R⁷;and R² and R³ are each independently —OR⁵, for example, R² and R³ areeach independently —OH. In some of the above embodiments, eachoccurrence of R⁷ is independently selected from halogen, —CF₃, —OCF₃,—C1-C4 aliphatic (e.g., —C1-C4 alkyl), and —O(C1-C4 aliphatic) (e.g.,—O(C1-C4 alkyl)). In some of the above embodiments, when Y¹ is a—CH₂—C(O)—, wherein the carbonyl group of Y¹ is connected directly to A,A is not:

(i) unsubstituted phenyl; or

(ii) phenyl substituted with halo (e.g., F— and Cl—) or lower alkyl(e.g., C1-C7 alkyl)

Examples of particular compounds of the present disclosure include:

or pharmaceutically acceptable salts thereof. In certain embodiments,the pharmaceutically acceptable salt is a sodium salt.

Examples of particular compounds of the present disclosure include:

or pharmaceutically acceptable salts thereof. In certain embodiments,the pharmaceutically acceptable salt is a sodium salt.

In another embodiment, the present disclosure provides a pharmaceuticalcomposition comprising a pharmaceutically acceptable carrier and acompound of formula I (including formula I-A and formula I-B) or formulaII or pharmaceutically acceptable salt form thereof. In yet otherembodiments, the present disclosure provides a pharmaceuticalcomposition comprising a pharmaceutically acceptable carrier and any oneof compounds 1-45 or a pharmaceutically acceptable salt form thereof. Incertain embodiments, the present disclosure provides a pharmaceuticalcomposition comprising a pharmaceutically acceptable carrier and thefollowing compound or a pharmaceutically acceptable salt form thereof:

In another embodiment, the present disclosure provides a compounddescribed herein in isolated form, i.e., an isolated compound. Exemplaryembodiments are an isolated compound of formula I (including formula I-Aand formula I-B), an isolated compound of formula II, or any ofcompounds 1-45 in isolated form. The term “isolated” refers to materialthat is removed from its original environment (e.g., the naturalenvironment if it is naturally occurring or a synthetic mixture if thematerial is synthesized in vitro or ex vivo). The isolated compound isdesirably substantially pure, such as having a purity of at least about80%, 85%, 90%, 95%, or 99% by weight.

It is understood that the compounds of the invention (e.g., compounds1-45) including pharmaceutically acceptable salts thereof andpharmaceutical compositions comprising at least one of the compounds ofthe invention (e.g., compounds 1-45), can be used in each of the methodsdescribed hereineblow.

C. General Synthetic Methodology

The compounds of this disclosure may be prepared in general by methodsknown to those skilled in the art. Scheme 1 below illustrates a generalsynthetic route to the compounds of the present disclosure. Otherequivalent schemes, which will be readily apparent to the ordinaryskilled organic chemist, may alternatively be used to synthesize variousportions of the molecules as illustrated by the general scheme below.

In another embodiment, the present disclosure provides a pharmaceuticalcomposition comprising a pharmaceutically acceptable carrier and acompound described herein, such as a compound of formula I or II.

The disclosure contemplates that any one or more of the foregoingaspects and embodiments (including compounds of all of the precedingformulae as well as their salts and prodrugs) can be combined with eachother and/or with any of the embodiments or features provided below.

D. Exemplary Uses

1. Neuronal Diseases/Disorders

In certain aspects, the compounds and compositions as described hereincan be used to treat patients suffering from P₂Y₆ receptor-relatedconditions or conditions that can be ameliorated by agonizing P₂Y₆receptor activity, such as neurodegenerative diseases, and traumatic ormechanical injury to the central nervous system (CNS), spinal cord orperipheral nervous system (PNS). Any of the compounds and compositions,including pharmaceutical compositions, of the disclosure can be used invitro or in vivo, including in any one or more of the in vivo or invitro methods described herein. For example, any of the compounds andcompositions of the disclosure can be used in vitro or in vivo, forexample to modulate P₂Y₆ receptor activity and/or to treat any one ormore of the diseases or conditions described herein. Moreover, any ofthe compounds and compositions of the disclosure can be used in vitro orin vivo, for example to modulate P₂Y₆ receptor activity and/or toinhibit release and/or reduce elevated levels of pro-inflammatorycytokines and/or to decrease levels of pro-inflammatory cytokines, suchas in plasma or secreted from cells. The disclosure contemplates thatany of the compounds of the disclosure, including salts thereof, may beused in any of the in vitro or in vivo methods described herein, such asto treat any one or more of the diseseas or conditions described herein.Similarly any of the compounds of the disclosure may be used in vitro,such as to modulate P₂Y₆ receptor activity. Moreover, any of thecompounds of the disclosure may be formulated as a pharmaceuticalcomposition comprising a compound and one or more acceptable carriersand/or excipients. Compositions, such as pharmaceutical compositions,may be used in any of the in vitro or in vivo methods described herein,such has to treat any one or more of the diseases or conditionsdescribed herein.

Accordingly, the disclosure contemplates methods of treating (decreasingthe frequency or severity of or otherwise alleviating one or moresymptoms of the condition) a subject in need thereof (e.g., a subjecthaving any of the conditions described herein, including any of theneurodegenerative or neuronal conditions described herein) byadministering a compound of the disclosure. Many of these, as well asother conditions described herein, are characterized by a level ofcognitive impairment and/or some decrease or loss of cognitive function.Cognitive function and cognitive impairment are used as understood inthe art. For example, cognitive function generally refers to the mentalprocesses by which one becomes aware of, perceives, or comprehendsideas. Cognitive function involves all aspects of perception, thinking,learning, reasoning, memory, awareness, and capacity for judgment.Cognitive impairment generally refers to conditions or symptomsinvolving problems with thought processes. This may manifest itself inone or more symptoms indicating a decrease in cognitive function, suchas impairment or decrease of higher reasoning skills, forgetfulness,impairments to memory, learning disabilities, concentrationdifficulties, decreased intelligence, and other reductions in mentalfunctions.

Neurodegenerative disease typically involves reductions in the mass andvolume of the human brain, which may be due to the atrophy and/or deathof brain cells, which are far more profound than those in a healthyperson that are attributable to aging. Neurodegenerative diseases canevolve gradually, after a long period of normal brain function, due toprogressive degeneration (e.g., nerve cell dysfunction and death) ofspecific brain regions. Alternatively, neurodegenerative diseases canhave a quick onset, such as those associated with trauma or toxins. Theactual onset of brain degeneration may precede clinical expression bymany years. Examples of neurodegenerative diseases include, but are notlimited to, Alzheimer's disease (AD), Parkinson's disease (PD),Huntington's disease (HD), amyotrophic lateral sclerosis (ALS; LouGehrig's disease), diffuse Lewy body disease, chorea-acanthocytosis,primary lateral sclerosis, ocular diseases (ocular neuritis),chemotherapy-induced neuropathies (e.g., from vincristine, paclitaxel,bortezomib), diabetes-induced neuropathies and Friedreich's ataxia. P₂Y₆receptor-modulating compounds of the present disclosure can be used totreat these disorders and others as described below.

AD is a CNS disorder that results in memory loss, unusual behavior,personality changes, and a decline in thinking abilities. These lossesare related to the death of specific types of brain cells and thebreakdown of connections and their supporting network (e.g. glial cells)between them. The earliest symptoms include loss of recent memory,faulty judgment, and changes in personality. Without being bound bytheory, these changes in the brain and symptoms associated withcognitive impairment, including memory and learning impairment, arecaused, in whole or in part, by accumulation of beta amyloid and theresulting deposition of amyloid plaques. PD is a CNS disorder thatresults in uncontrolled body movements, rigidity, tremor, anddyskinesia, and is associated with the death of brain cells in an areaof the brain that produces dopamine. ALS (motor neuron disease) is a CNSdisorder that attacks the motor neurons, components of the CNS thatconnect the brain to the skeletal muscles.

HD is another neurodegenerative disease that causes uncontrolledmovements, loss of intellectual faculties, and emotional disturbance.Tay-Sachs disease and Sandhoff disease are glycolipid storage diseaseswhere GM2 ganglioside and related glycolipids substrates forβ-hexosaminidase accumulate in the nervous system and trigger acuteneurodegeneration.

It is well-known that apoptosis plays a role in AIDS pathogenesis in theimmune system. However, HIV-1 also induces neurological disease, whichcan be treated with P₂Y₆ receptor-modulating compounds of thedisclosure.

Neuronal loss is also a salient feature of prion diseases, such asCreutzfeldt-Jakob disease in human, BSE in cattle (mad cow disease),Scrapie Disease in sheep and goats, and feline spongiform encephalopathy(FSE) in cats. P₂Y₆ receptor-modulating compounds as described herein,may be useful for treating or preventing neuronal loss due to theseprion diseases.

In another embodiment, the compounds as described herein may be used totreat or prevent any disease or disorder involving axonopathy. Distalaxonopathy is a type of peripheral neuropathy that results from somemetabolic or toxic derangement of peripheral nervous system (PNS)neurons. It is the most common response of nerves to metabolic or toxicdisturbances, and as such may be caused by metabolic diseases such asdiabetes, renal failure, deficiency syndromes such as malnutrition andalcoholism, or the effects of toxins or drugs. Those with distalaxonopathies usually present with symmetrical glove-stockingsensori-motor disturbances. Deep tendon reflexes and autonomic nervoussystem (ANS) functions are also lost or diminished in affected areas.

Diabetic neuropathies are neuropathic disorders that are associated withdiabetes mellitus. Relatively common conditions which may be associatedwith diabetic neuropathy include third nerve palsy; mononeuropathy;mononeuritis multiplex; diabetic amyotrophy; a painful polyneuropathy;autonomic neuropathy; and thoracoabdominal neuropathy.

Peripheral neuropathy is the medical term for damage to nerves of theperipheral nervous system, which may be caused either by diseases of thenerve or from the side-effects of systemic illness. Major causes ofperipheral neuropathy include seizures, nutritional deficiencies, andHIV, though diabetes is the most likely cause.

In an exemplary embodiment, a P₂Y₆ receptor-modulating compound asdescribed herein may be used to treat or prevent multiple sclerosis(MS), including relapsing MS and monosymptomatic MS, and otherdemyelinating conditions, such as, for example, chronic inflammatorydemyelinating polyneuropathy (CIDP), or symptoms associated therewith.

In yet another embodiment, compounds of the present disclosure may beused to treat trauma to the nerves, including, trauma due to disease,injury (including surgical intervention), or environmental trauma (e.g.,neurotoxins, alcoholism, etc.). In certain embodiments, compounds of thepresent disclosure may be used to treat traumatic brain injury, such asto improve cognitive function in a subject suffering from a traumaticbrain injury. Without being bound by theory, there is often an increasein beta amyloid observed following traumatic brain injuries. The presentdisclosure provides methods suitable for enhancing clearance of betaamyloid or otherwise reducing beta amyloid and/or plaque burden in asubject.

Compounds of the present disclosure may also be useful to prevent,treat, and alleviate symptoms of various PNS disorders. The term“peripheral neuropathy” encompasses a wide range of disorders in whichthe nerves outside of the brain and spinal cord—peripheral nerves—havebeen damaged. Peripheral neuropathy may also be referred to asperipheral neuritis, or if many nerves are involved, the termspolyneuropathy or polyneuritis may be used.

PNS diseases treatable with P₂Y₆ receptor-modulating compounds asdescribed herein, include: diabetes, leprosy, Charcot-Marie-Toothdisease, Guillain-Barré syndrome and Brachial Plexus Neuropathies(diseases of the cervical and first thoracic roots, nerve trunks, cords,and peripheral nerve components of the brachial plexus).

In another embodiment, compounds of the present disclosure may be usedto treat or prevent a polyglutamine disease. Exemplary polyglutaminediseases include Spinobulbar muscular atrophy (Kennedy disease),Huntington's Disease (HD), Dentatorubral-pallidoluysian atrophy (HawRiver syndrome), Spinocerebellar ataxia type 1, Spinocerebellar ataxiatype 2, Spinocerebellar ataxia type 3 (Machado-Joseph disease),Spinocerebellar ataxia type 6, Spinocerebellar ataxia type 7, andSpinocerebellar ataxia type 17.

In certain embodiments, the disclosure provides a method to treat acentral nervous system cell to prevent damage in response to a decreasein blood flow to the cell. Typically the severity of damage that may beprevented will depend in large part on the degree of reduction in bloodflow to the cell and the duration of the reduction. In some embodiments,apoptotic or necrotic cell death may be prevented. In still a furtherembodiment, ischemic-mediated damage, such as cytoxic edema or centralnervous system tissue anoxemia, may be prevented. In each embodiment,the central nervous system cell may be a spinal cell or a brain cell.

Another aspect encompasses administrating a compound as described hereinto a subject to treat a central nervous system ischemic condition. Anumber of central nervous system ischemic conditions may be treated bythe compounds described herein.

In some embodiments, the ischemic condition is a stroke that results inany type of ischemic central nervous system damage, such as apoptotic ornecrotic cell death, cytoxic edema or central nervous system tissueanoxia. The stroke may impact any area of the brain or be caused by anyetiology commonly known to result in the occurrence of a stroke. In onealternative of this embodiment, the stroke is a brain stem stroke. Inanother alternative of this embodiment, the stroke is a cerebellarstroke. In still another embodiment, the stroke is an embolic stroke. Inyet another alternative, the stroke may be a hemorrhagic stroke. In afurther embodiment, the stroke is a thrombotic stroke.

In yet another aspect, compounds of the disclosure may be administeredto reduce infarct size of the ischemic core following a central nervoussystem ischemic condition. Moreover, compounds of the present disclosuremay also be beneficially administered to reduce the size of the ischemicpenumbra or transitional zone following a central nervous systemischemic condition.

In some embodiments, a combination drug regimen may include drugs orcompounds for the treatment or prevention of neurodegenerative disordersor secondary conditions associated with these conditions. Thus, acombination drug regimen may include one or more compounds as describedherein and one or more anti-neurodegeneration agents.

In a particular embodiment, the disclosure provides methods for doingone or more of decreasing plaque burden, improving cognitive function,decreasing or delaying cognitive impairment, or improving hippocampallong term potentiation by administering to a subject in need thereof acompound of the disclosure. These methods may also be used for one ormore of enhancing beta amyloid clearance, increasing synapticplasticity, or improving or restoring memory. The foregoing areexemplary of beneficial results that would help alleviate (e.g., treat)one or more symptoms of conditions associated with cognitive impairment.Exemplary conditions include AD, traumatic brain injury, and DownSyndrome, as well as other neurological and neurodegenerative diseases.Moreover, the disclosure contemplates the alleviation of symptoms inconditions and scenarios associated with milder forms of cognitiveimpairment, such as age-related dementia, mild cognitive impairment, andeven to improve memory and cognitive function that typically declines,even in relatively healthy individuals, as part of the normal agingprocess. Exemplary such agonists are described herein, and thedisclosure contemplates that any such compounds can be used in thetreatment of any of the conditions described herein. Regardless ofwhether one of the agonists described herein are used or whether anotheragonist is used, the disclosure contemplates that the agonist may beformulated in a pharmaceutically acceptable carrier and administered byany suitable route of administration. These methods are of particularuse when the subject in need thereof has Alzheimer's disease. It isunderstood by those of skill in the art that definitive diagnosis ofAlzheimer's disease is difficult and may require postmortem examination.Thus, in this context and in the context of the present disclosure,having Alzheimer's disease is used to refer to subjects who have beendiagnosed with Alzheimer's disease or who are suspected by a physicianof having Alzheimer's disease. However, these methods are also ofparticular use when the subject in need thereof has any other conditionassociated with cognitive impairment, for example, a condition in whichthe impairment is accompanied with an increase in beta amyloid, adecrease in the rate of beta amyloid clearance, and/or an increase inamyloid plaque deposition.

Cognitive function and cognitive impairment may be readily evaluatedusing tests well known in the art. Performance in these tests can becompared over time to determine whether a treated subject is improvingor whether further decline has stopped or slowed, relative to theprevious rate of decline of that patient or compared to an average rateof decline. Exemplary tests used in animal studies are provided in, forexample, Animal Models of Cognitive Impairment, Levin E D, Buccafusco JJ, editors. Boca Raton (Fla.): CRC Press; 2006. Tests of cognitivefunction, including memory and learning for evaluating human patientsare well known in the art and regularly used to evaluate and monitorsubjects having or suspected of having cognitive disorders such as AD.Even in healthy individuals, these and other standard tests of cognitivefunction can be readily used to evaluate beneficial affects over time.

Compounds of the disclosure are also useful in the treatment ofParkinson's disease. For example, compounds of the disclosure can beused to improve the motor impairments symptomatic of Parkinson'sdisease. Moreover, compounds of the disclosure are useful for treatingthe memory impairment symptomatic of Parkinson's disease. Without beingbound by theory, impairment of microglial phagocytosis is thought to bea mechanism of action underlying accumulation of alpha synuclein and theformation of Lewy bodies (and resulting neurodegeneration) inParkinson's disease. Compounds of the disclosure may be used to increaseclearance or otherwise decrease extracellular alpha-synuclein, todecrease intracellular accumulation of alpha-synuclein, and/or todecrease or prevent the formation of Lewy bodies in a subject in needthereof. In certain embodiments, compounds of the disclosure enhancephagocytosis, such as microglial phagocytosis.

Compounds of the disclosure, including salts and prodrugs, may be testedin animal models of Parkinson's disease. Exemplary models include micethat over express α-synuclein, express human mutant forms ofα-synuclein, mice that express LRKK2 mutations, as well as mice treatedwith MTTP. Additional information regarding these animal models isreadily available from Jackson Laboratories (see also the websiteresearch.jax.org/grs/parkinsons.html), as well as in numerouspublications disclosing the use of these validated models.

2. Down Syndrome

Compounds of the present disclosure may also be useful to prevent,treat, and alleviate symptoms of Down Syndrome (DS). Down Syndrome (DS)is a genetic condition characterized by trisomy of chromosome 21. DS isnamed after Dr. John Langdon Down, an English physician who firstdescribed the characteristics of DS in 1866. It was not until 1959 thatJerome Leieune and Patricia Jacobs independently first determined thecause to be trisomy of the 21st chromosome.

In recent years, it has become evident that there is relationshipbetween Alzheimer's Disease (AD) and DS. Specifically, the production ofexcessive beta amyloid plaques and amyloid angiopathy occurs in both DSand Alzheimer's Disease (AD) (Delabar et al. (1987) “Beta amyloid genetriplication in Alzheimer's disease and karyotypically normal DownSyndrome. Science 235: 1390-1392). Without being bound by theory, giventhat both AD and Down Syndrome are characterized by both beta amyloidplaques and cognitive impairment, methods and compositions that decreaseplaque burden and/or enhance beta amyloid clearance are useful fortreating AD and Down Syndrome (e.g., providing a beneficial effectand/or decreasing one or more symptoms of AD or Down Syndrome).Exemplary beneficial effects include, but are not limited to, improvingcognitive function, decreasing cognitive impairment, decreasing plaqueburden, enhancing beta amyloid clearance, improving memory, and thelike.

3. Pain

In certain aspects, the compounds as described herein can be used totreat patients having pain. Pain is a complex physiological process thatinvolves a number of sensory and neural mechanisms. Compounds to be usedaccording to the present disclosure are suitable for administration to asubject for treatment (including prevention and/or alleviation) ofchronic and/or acute pain, in particular non-inflammatorymusculoskeletal pain such as back pain, fibromyalgia and myofascialpain, more particularly for reduction of the associated muscularhyperalgesia or muscular allodynia. Nonlimiting examples of types ofpain that can be treated by the compounds, compositions and methods ofthe present disclosure include chronic conditions such asmusculoskeletal pain, including fibromyalgia, myofascial pain, backpain, pain during menstruation, pain during osteoarthritis, pain duringrheumatoid arthritis, pain during gastrointestinal inflammation, painduring inflammation of the heart muscle, pain during multiple sclerosis,pain during neuritis, pain during AIDS, pain during chemotherapy, tumorpain, headache, CPS (chronic pain syndrome), central pain, neuropathicpain such as trigeminal neuralgia, shingles, stamp pain, phantom limbpain, temporomandibular joint disorder, nerve injury, migraine,post-herpetic neuralgia, neuropathic pain encountered as a consequenceof injuries, amputation infections, metabolic disorders or degenerativediseases of the nervous system, neuropathic pain associated withdiabetes, pseudesthesia, hypothyroidism, uremia, vitamin deficiency oralcoholism; and acute pain such as pain after injuries, postoperativepain, pain during acute gout or pain during operations, such as jawsurgery.

Acute pain is typically a physiological signal indicating a potential oractual injury. Chronic pain can be somatogenic (organic) or psychogenic.Chronic pain is frequently accompanied or followed by vegetative signs,such as, for example, lassitude or sleep disturbance. Acute pain may betreated with compounds as described herein.

Somatogenic pain may be of nociceptive, inflammatory or neuropathicorigin. Nociceptive pain is related to activation of somatic or visceralpain-sensitive nerve fibers, typically by physical or chemical injury totissues. Inflammatory pain results from inflammation, for example aninflammatory response of living tissues to any stimulus includinginjury, infection or irritation. Neuropathic pain results fromdysfunction in the nervous system. Neuropathic pain is believed to besustained by aberrant somatosensory mechanisms in the peripheral nervoussystem, the central nervous system (CNS), or both. According to oneaspect of the disclosure, somatogenic pain may be treated by compoundsas described herein.

Non-inflammatory musculoskeletal pain is a particular form of chronicpain that is generally not traced to a specific structural orinflammatory cause and that generally does not appear to be induced bytissue damage and macrophage infiltration (resulting in edema) as occursin a classical immune system response. Although non-inflammatorymusculoskeletal pain is believed to result from peripheral and/orcentral sensitization, the cause is not presently fully understood. Itis often associated with physical or mental stress, lack of adequate orrestful sleep, or exposure to cold or damp. Non-inflammatorymusculoskeletal pain is also believed to be associated with orprecipitated by systemic disorders such as viral or other infections.Examples of non-inflammatory musculoskeletal pain include neck andshoulder pain and spasms, low back pain, and achy chest or thighmuscles, which may be treated by a compound of the present disclosure.Non-inflammatory musculoskeletal pain may be generalized or localized.

According to a further aspect of the disclosure, a compound as describedherein may be administered to a subject to treat fibromyalgia syndrome(FMS) and myofascial pain syndrome (MPS). FMS and MPS are medicalconditions characterized by fibromyalgia and myofascial painrespectively, which are two types of non-inflammatory musculoskeletalpain. FMS is a complex syndrome associated with significant impairmentof quality of life and can result in substantial financial costs.Fibromyalgia is a systemic process that typically causes tender points(local tender areas in normal-appearing tissues) in particular areas ofthe body and is frequently associated with a poor sleep pattern and/orstressful environment. Diagnosis of fibromyalgia is typically based on ahistory of widespread pain (e.g., bilateral, upper and lower body,and/or spinal pain), and presence of excessive tenderness on applyingpressure to a number of (sometimes more precisely defined as at least 11out of 18) specific muscle-tender sites. FMS is typically a chronicsyndrome that causes pain and stiffness throughout the tissues thatsupport and move the bones and joints. Myofascial pain syndrome (MPS) isa chronic non-degenerative, non-inflammatory musculoskeletal conditionoften associated with spasm or pain in the masticatory muscles. Distinctareas within muscles or their delicate connective tissue coverings(fascia) become abnormally thickened or tight. When the myofascialtissues tighten and lose their elasticity, the ability ofneurotransmitters to send and receive messages between the brain andbody is disrupted. Specific discrete areas of muscle may be tender whenfirm fingertip pressure is applied; these areas are called tender ortrigger points. Symptoms of MPS include muscle stiffness and aching andsharp shooting pains or tingling and numbness in areas distant from atrigger point. The discomfort may cause sleep disturbance, fatigue anddepression. Most commonly trigger points are in the jaw(temporomandibular) region, neck, back or buttocks. Myofascial paindiffers from fibromyalgia: MPS and FMS are two separate entities, eachhaving its own pathology, but sharing the muscle as a common pathway ofpain. Myofascial pain is typically a more localized or regional (alongthe muscle and surrounding fascia tissues) pain process that is oftenassociated with trigger point tenderness. Myofascial pain can be treatedby a variety of methods (sometimes in combination) including stretching,ultrasound, ice sprays with stretching, exercises, and injections ofanesthetic.

A further non-inflammatory musculoskeletal pain condition is back pain,notably low back pain, which may also be treated with a compound of thepresent disclosure. This condition may also be treating by administeringa compound of the present disclosure to a subject in need thereof. Backpain is a common musculoskeletal symptom that may be either acute orchronic. It may be caused by a variety of diseases and disorders thataffect the lumbar spine. Low back pain is often accompanied by sciatica,which is pain that involves the sciatic nerve and is felt in the lowerback, the buttocks, and the backs of the thighs.

4. Glaucoma and Intraocular Pressure

In another aspect, the disclosure provides for methods of treatingglaucoma in a subject in need thereof. Compounds of the disclosure, suchas any of the compounds described herein, may be used to treat glaucoma.For example, compounds of the disclosure may be used to decreaseintraocular pressure (IOP), such as the elevated intraocular pressureobserved in most cases of glaucoma. Also provided are methods fortreating ocular hypertension in a subject in need thereof. Without beingbound by theory, compounds of the disclosure may be used to reduce IOP,thereby treating ocular hypertension. For any of the foregoing, thedisclosure contemplates administering an effective amount of a P₂Y₆receptor agonist, such as any of the agonists described herein, to asubject in need thereof to decrease intraocular pressure, such aselevated intraocular pressure and/or to treat glaucoma (e.g., improve,or stop or slow the progression of one or more symptoms of thecondition).

Glaucoma refers to a group of eye conditions that lead to damage to theoptic nerve. This nerve carries visual information from the eye to thebrain. In most cases, damage to the optic nerve is due to increasedpressure in the eye, also known as intraocular pressure (IOP). Overtime, the elevated intraocular pressure and optic nerve damage leads tovisual field loss, and may result in blindness. Ocular hypertension isintraocular pressure higher than normal in the absence of optic nervedamage or visual field loss. Currently, ophthalmologists generallydefine normal intraocular pressure as from 10 mmHg and 21 mmHg, andintraocular pressure above 21 mmHg is considered ocular hypertension orelevated intraocular pressure. Ocular hypertension is considered asignificant risk factor for developing glaucoma, and thus, patients withocular hypertension should be closely monitored for glaucoma.

Glaucoma is the second-most common cause of blindness in the UnitedStates. The nerve damage involves loss of retinal ganglion cells in acharacteristic pattern. The many different subtypes of glaucoma can allbe considered to be a type of optic neuropathy. Raised intraocularpressure (above 21 mmHg or 2.8 kPa) is the most important and onlymodifiable risk factor for glaucoma. However, some patients may havehigh eye pressure for years and never develop damage, while others candevelop nerve damage at a relatively low pressure. Untreated glaucomacan lead to permanent damage of the optic nerve and resultant visualfield loss, which over time can progress to blindness.

The two main types of glaucoma, each of which are marked by elevatedintraocular pressure, are open-angle and angle-closure. Open-angle andangle-closure glaucoma also include the following variants: (i)secondary glaucoma; (ii) pigmentary glaucoma; (iii) pseudoexfoliativeglaucoma; (iv) traumatic glaucoma; (v) neovascular glaucoma; and (vi)irido corneal endothelial syndrome (ICE).

Open-angle glaucoma, the most common form of glaucoma, accounts for atleast 90% of all glaucoma cases. Open-angle glaucoma is also calledprimary or chronic glaucoma and generally has the followingcharacteristics: (i) caused by the slow clogging of the drainage canals,resulting in increased eye pressure; (ii) has a wide and open anglebetween the iris and cornea; and (iii) develops slowly and is a lifelongcondition. Angle-closure glaucoma, a less common form of glaucoma, isalso called acute glaucoma or narrow-angle glaucoma. Unlike open-angleglaucoma, angle-closure glaucoma is a result of the angle between theiris and cornea closing, and angle-closure glaucoma generally has thefollowing characteristics: (i) caused by blocked drainage canals,resulting in a sudden rise in intraocular pressure; (ii) has a closed ornarrow angle between the iris and cornea; (iii) develops very quickly;and (iv) demands immediate medical attention.

The disclosure contemplates methods of treating open-angle and/orangle-closure glaucoma, including methods of treating variants ofopen-angle and/or angle-closure glaucoma. In certain embodiments,administration of a compound of the disclosure to a patient havingopen-angle and/or angle-closure glaucoma decreases intraocular pressure,thereby treating the glaucoma in the patient. In certain embodiments,reducing intraocular pressure slows or stops further damage to the opticnerve (e.g., prevents occurrence of further damage to the optic nerve).In certain embodiments, reducing intraocular pressure slows or stopsfurther loss of or damage to vision or the visual field. Moreover, thedisclosure contemplates methods of treating optic neuropathy in apatient in need thereof by administering an effective amount of acompound of the disclosure.

In addition to open-angle and angle-closure glaucoma, an additional raretype of glaucoma is congenital glaucoma. In certain embodiments, thedisclosure contemplates methods of treating congenital glaucoma in asubject in need thereof. This type of glaucoma occurs in babies whenthere is incorrect or incomplete development of the eye's drainagecanals during the prenatal period.

A fourth type of glaucoma is referred to as secondary glaucoma.Secondary glaucoma occurs as a consequence of trauma, systemic disease,or as a side-effect of certain drugs (e.g., corticosteroids). In certainembodiments, the disclosure contemplates methods of treating secondaryglaucoma in a subject in need thereof. Systemic diseases that may leadto or exacerbate glaucoma include hypertension and diabetes.

Additionally, although most glaucoma is characterized by elevatedintraocular pressure which leads to damage of the optic nerve, there arecases of glaucoma referred to as low-tension or normal-pressureglaucoma. In these cases, the optic nerve is damaged despite the factthat eye pressure is not very high. In certain embodiments, thedisclosure contemplates methods of treating low-tension or normalpressure glaucoma. In certain embodiments, glaucoma treated using themethods of the disclosure is characterized by elevated intraocularpressure and/or ocular hypertension (e.g., the glaucoma is notlow-tension or normal pressure glaucoma).

In addition to elevated intraocular pressure, which results in damage tothe optic nerve, the various types of glaucoma are characterized byparticular symptoms. The disclosure contemplates that administration ofthe compounds of this disclosure may be used to alleviate one or moresymptoms of glaucoma, including to alleviate one or more symptoms of anyof the particular types of glaucoma described herein.

In open-angle glaucoma, there are actually few overt symptoms. Patientshave elevated intraocular pressure or, at least, periods of elevatedintraocular pressure. However, the intraocular pressure slowly damagesthe optic nerve, and thus, vision loss is slow and not typicallyaccompanied by pain. In fact, noticeable vision loss, which typicallymanifests as slow loss of peripheral vision leading to tunnel vision, isa symptom of relatively advanced and severe disease. Ultimately,open-angle glaucoma can lead to blindness.

In angle-closure glaucoma, patients do experience one or more of thefollowing symptoms, and these symptoms may come and go or steadilybecome worse. Exemplary symptoms include sudden, severe pain (typicallyin only one eye), decreased or cloudy vision (also known as “steamy”vision), nausea, vomiting, rainbow-like halos around lights, red eye,and the sensation that the eye is swollen.

In congenital glaucoma, the symptoms are usually noticed when the childis a few months old. Exemplary symptoms include one or more of thefollowing: cloudiness of the front of the eye, enlargement of one orboth eyes, red eye, sensitivity to light, and excessive tearing.

There are currently several tests that are used to measure intraocularpressure, to detect elevated intraocular pressure and to diagnoseglaucoma. In certain embodiments, one or more of these tests are used todiagnose glaucoma and/or intraocular hypertension prior to initiation oftreatment with a compound of the disclosure. Exemplary tests that can beused, alone or in combination, include tonometry, gonioscopy, opticnerve imaging, slit lamp examination, examination of the retina, visualacuity measurements, and visual field measurements. These tests can alsobe used to monitor a patient after initiation of treatment. For example,these tests can be used to determine whether treatment has slowed orstopped the progress of the disease, has decreased elevated intraocularpressure (e.g., restored normal intraocular pressure), and whether thepatient's vision has improved or ceased further deterioration.

In addition, the disclosure provides methods of decreasing elevatedintraocular pressure in a subject in need thereof. Suitable subjectsinclude, as discussed in detail above, subjects having glaucoma (any ofthe forms of glaucoma described herein) or subjects with ocularhypertension. Decreasing intraocular pressure, such as elevatedintraocular pressure in these subjects (e.g., such as by administeringan effective amount of a compound of the disclosure, including any ofthe compounds described herein), such as human patients, helpsameliorate one or more symptoms of the condition, helps slow or stopdamage to the optic nerve and to vision, and may even permit improvementin the patient's condition—particularly in cases where significantdamage has not yet occurred. Given that elevated intraocular pressure inocular hypertension is a major risk factor for developing glaucoma,decreasing elevated IOP in such patients may help decrease the patient'srisk of developing glaucoma.

Compounds of the disclosure, including salts and prodrugs, may be testedin animal models of glaucoma and ocular hypertension. Exemplary modelsare known in the art and, for example, described in Bouhenni et al.,Journal of Biomedicine and Biotechnology, Volumne 2012, Article ID692609, 11 pages, doi: 10.1155/2012/692609.

Agonists of the disclosure may be administered using any suitable routeof administration described herein, including oral, intravenous, orlocal administration to the eye (e.g., eye drops, injection into theeye, or implantation of a drug eluting device).

In other embodiments, the disclosure provides methods of decreasingintraocular pressure (e.g., decreasing elevated intraocular pressure) ina subject in need thereof, wherein the subject in need thereof has acondition other than or in addition to glaucoma. Exemplary conditionscaused or exacerbated by elevated IOP which may be treated include:Reese-Ellsworth syndrome, hydrophthalmos, and ophthalmic zoster.

5. Inflammatory Conditions

In another aspect, the compounds, salts and/or prodrugs thereof, andcompositions as described herein can be used to treat patients sufferingfrom P₂Y₆ receptor-related conditions or conditions that can beameliorated by modulating, for example, agonizing P₂Y₆ receptoractivity, such as an inflammatory condition. Accordingly, the disclosureprovides methods of treating an inflammatory condition in a subject inneed thereof. Compounds of the disclosure, such as any of the compoundsor salts or prodrugs described herein, may be used to treat aninflammatory condition. As used herein, an inflammatory condition is adisease or condition characterized, in whole or in part, by inflammationor an inflammatory response in the patient. Typically, one or more ofthe symptoms of the inflammatory disease or condition is caused orexacerbated by an inappropriate, misregulated, or overactiveinflammatory response. Inflammatory diseases or conditions may bechronic or acute. In certain embodiments, the inflammatory disease orcondition is an autoimmune disorder. In certain embodiments, compoundsof the disclosure are used to decrease inflammation, to decreaseexpression of one or more inflammatory cytokines, and/or to decrease anoveractive inflammatory response in a subject having an inflammatorycondition. Thus, the disclosure provides a method of decreasinginflammation, a method of decreasing expression of one or moreinflammatory cytokines, and/or a method of decreasing an overactiveinflammatory response in a subject in need thereof.

Inflammatory conditions treatable using the compounds of the disclosuremay be characterized, for example, based on the primary tissue affected,the mechanism of action underlying the condition, or the portion of theimmune system that is misregulated or overactive. Examples ofinflammatory conditions, as well categories of diseases and conditionsare provided herein. The disclosure contemplates methods of treating(e.g., such as by decreasing inflammation, decreasing expression of oneor more inflammatory cytokines, and/or decreasing an overactiveinflammatory response) inflammatory conditions, generally, as well asmethods of treating any of the categories of conditions or any of thespecific conditions described herein.

In certain embodiments, examples of inflammatory conditions that may betreated include inflammation of the lungs, joints, connective tissue,eyes, nose, bowel, kidney, liver, skin, central nervous system, vascularsystem, heart, or adipose tissue. In certain embodiments, inflammatoryconditions which may be treated include inflammation due to theinfiltration of leukocytes or other immune effector cells into affectedtissue. In certain embodiments, inflammatory conditions which may betreated include inflammation mediated by IgE antibodies. Other relevantexamples of inflammatory conditions which may be treated by the presentdisclosure include inflammation caused by infectious agents, includingbut not limited to viruses, bacteria, fungi, and parasites. In certainembodiments, the inflammatory condition that is treated is an allergicreaction. In certain embodiments, the inflammatory condition is anautoimmune disease. The disclosure contemplates that some inflammatoryconditions involve inflammation in multiple tissues. Moreover, thedisclosure contemplates that some inflammatory conditions may fall intomultiple categories. For example, a condition may be described andcategorized as an autoimmune condition and/or it may also be describedand categorized based on the primary tissue(s) affected (e.g., aninflammatory skin or joint condition). In certain embodiments, aninflammatory condition treatable according to the methods describedherein falls into more than one category of condition.

Inflammatory lung conditions include asthma, adult respiratory distresssyndrome, bronchitis, pulmonary inflammation, pulmonary fibrosis, andcystic fibrosis (which may additionally or alternatively involve thegastro-intestinal tract or other tissue(s)). In certain embodiments, thepulmonary inflammation is allergen induced pulmonary inflammation. Incertain embodiments, the disclosure provides methods of treating aninflammatory lung condition in a patient in need thereof (e.g., apatient having an inflammatory lung condition) by administering aneffective amount a compound, salt, or prodrug of the disclosure. Incertain embodiments, treating an inflammatory lung condition comprisesdecreasing inflammation in the lung in the patient, decreasingmisregulation of inflammatory cytokines in the patient, and/ordecreasing one or more symptoms of the inflammatory lung condition inthe subject. By way of example, symptoms of the inflammatory lungcondition that may be improved, locally or systemically, by decreasinginflammation or the inflammatory response include, but are not limitedto: oxygen saturation (patients have improved oxygen saturationfollowing treatment), ease of breathing (patients experience greaterease when breathing and a decrease in labored breather), reliance onexternal oxygen (patient reliance on external oxygen supplementation isdecreased), and reliance on inhalers or nebulizers (patient reliance onother drugs is decreased). Improvement in a patient (e.g., decrease insymptoms) may be measured directly by assessing inflammation or scarringin the lung or by evaluating cytokine expression in lung fluids.Improvement can also be assessed by evaluating improvement in patientactivity levels, walking distance and speed, and decreased reliance onoxygen supplementation.

Inflammatory joint conditions include rheumatoid arthritis, rheumatoidspondylitis, juvenile rheumatoid arthritis, osteoarthritis, goutyarthritis and other arthritic conditions. In certain embodiments, theinflammatory joint condition is rheumatoid arthritis or psoriaticarthritis. In certain embodiments, the disclosure provides methods oftreating an inflammatory joint condition in a patient in need thereof,such as treating any of the foregoing conditions, by administering aneffective amount of a compound, salt and/or prodrug of the disclosure.In certain embodiments, treating an inflammatory joint conditioncomprises decreasing inflammation in the joints in the patient,decreasing circulating levels of one or more cytokines, for example,IL-4, IL-10 and/or IL-12, in plasma of the patient, decreasingmisregulation of inflammatory cytokines in the patient, and/ordecreasing one or more symptoms of the inflammatory joint condition inthe subject. By way of example, symptoms of the inflammatory jointcondition that may be improved by decreasing inflammation or theinflammatory response, locally and/or systemically, include, but are notlimited to: swelling in one or more joints, tenderness and/or pain inone or more joints, decreased mobility and/or use of one or more joints,impaired ability to perform daily tasks (e.g., ability to perform dailytasks including self care tasks is improved), and reliance on walkingassistance (patient reliance on a walker, cane, or wheel chair isdecreased). Improvement in patients (e.g., decrease in symptoms) may bemeasured directly by assessing inflammation in the joints or byevaluating cytokine expression in joint fluid. Improvement can also beassessed by evaluating improvement in patient activity levels andquality of life measures, walking distance and speed, range of motion,mobility, and decreased reliance on mobility aids. In certainembodiments, the inflammatory joint condition is also an autoimmunecondition, and the disclosure contemplates treating such condition.

Inflammatory eye conditions include uveitis (including iritis),conjunctivitis, scleritis, and keratoconjunctivitis sicca. In certainembodiments, the disclosure contemplates treating an inflammatory eyecondition in a patient in need thereof, including by administering acompound, salt and/or prodrug of the disclosure systemically or locallyto the eye, such as via eye drops.

Inflammatory bowel conditions include Crohn's disease, ulcerativecolitis, inflammatory bowel disease, inflammatory bowel syndrome, anddistal proctitis. In certain embodiments, the disclosure providesmethods of treating an inflammatory bowel condition in a patient in needthereof by administering an effective amount a compound, salt and/orprodrug of the disclosure. In certain embodiments, treating aninflammatory bowel condition comprises decreasing inflammation in thegastro-intestinal tract in the patient, decreasing misregulation ofinflammatory cytokines in the patient, decreasing the circulating levelsof one or more cytokines, for example, IL-4, IL-10 and/or IL-12, inplasma of the patient, and/or decreasing one or more symptoms of theinflammatory bowel condition in the subject. By way of example, symptomsof the inflammatory bowel condition that may be improved by decreasinginflammation or the inflammatory response, locally and/or systemically,include, but are not limited to: diarrhea, constipation, blotting, pain,flatulence, blood in stool, weight loss (treating stabilizes weightand/or prevents further weight loss; treatment helps promote improvednutrition and weight gain, where needed), malabsorption, andmalnutrition. Improvement in patients (e.g., decrease in symptoms) maybe measured directly by assessing inflammation in the gastrointestinaltract or by evaluating cytokine expression or levels of cytokines inplasma in patients. Improvement can also be assessed by evaluatingimprovement in any of the foregoing symptoms, evaluating patientself-reporting of quality of life and symptom reduction, evaluatingpatient weight and nutrition status. In certain embodiments, theinflammatory bowel condition being treated is also an autoimmunecondition, such as ulcerative colitis.

Inflammatory skin conditions include conditions associated with cellproliferation, such as psoriasis, eczema, and dermatitis (e. g.,eczematous dermatitides, topic and seborrheic dermatitis, allergic orirritant contact dermatitis, eczema craquelee, photoallergic dermatitis,phototoxicdermatitis, phytophotodermatitis, radiation dermatitis, andstasis dermatitis). Other inflammatory skin conditions include, but arenot limited to, ulcers and erosions resulting from trauma, burns,bullous disorders, or ischemia of the skin or mucous membranes, severalforms of ichthyoses, epidermolysis bullosae, hypertrophic scars,keloids, cutaneous changes of intrinsic aging, photo aging, frictionalblistering caused by mechanical shearing of the skin and cutaneousatrophy resulting from the topical use of corticosteroids. Additionalinflammatory skin conditions include inflammation of mucous membranes,such as cheilitis, nasal irritation, mucositis and vulvovaginitis. Otherinflammatory skin conditions include acne, rosacea, boils, carbuncles,pemphigus, cellulitis, Grover's disease, hidradenitis suppurativa, andlichen planus. In certain embodiments, the disclosure provides methodsof treating an inflammatory skin condition in a patient in need thereofby administering an effective amount a compound, salt and/or prodrug ofthe disclosure. In certain embodiments, treating an inflammatory skincondition comprises decreasing skin inflammation in the patient,decreasing misregulation of inflammatory cytokines in the patient,decreasing the circulating levels of one or more cytokines, for example,IL-4, IL-10 and/or IL-12, in plasma of the patient, and/or decreasingone or more symptoms of the inflammatory skin condition in the subject.By way of example, symptoms of the inflammatory skin condition that maybe improved by decreasing inflammation or the inflammatory response,locally and/or systemically, include, but are not limited to: skinswelling, redness, itching, flaking, blistering, bleeding, sensitivityto touch, and sensitivity to light or sun. Improvement in patients(e.g., decrease in symptoms) may be measured directly by assessinginflammation or by evaluating cytokine expression in patients.Improvement can also be assessed by evaluating improvement in any of theforegoing symptoms, or by evaluating patient self-reporting of qualityof life and symptom reduction. In certain embodiments, the inflammatoryskin condition is also an autoimmune condition, such as psoriasis. Thedisclosure provides methods of treating an inflammatory skin condition.

Inflammatory conditions of the endocrine system include, but are notlimited to, autoimmune thyroiditis (Hashimoto's disease), Type Idiabetes, inflammation in liver and adipose tissue associated with TypeII diabetes, and acute and chronic inflammation of the adrenal cortex.Inflammatory conditions of the cardiovascular system include, but arenot limited to, coronary infarct damage, peripheral vascular disease,myocarditis, vasculitis, revascularization of stenosis, atherosclerosis,and vascular disease associated with Type II diabetes. In certainembodiments, the disclosure provides methods of treating an inflammatoryendocrine condition or cardiovascular condition in a patient in needthereof by administering an effective amount a compound, salt and/orprodrug of the disclosure. In certain embodiments, treating aninflammatory endocrine condition or cardiovascular condition comprisesdecreasing inflammation in the patient, decreasing misregulation ofinflammatory cytokines in the patient, decreasing circulating levels ofone or more cytokines, for example, IL-4, IL-10 and/or IL-12, in plasmaof the patient, and/or decreasing one or more symptoms of theinflammatory endocrine condition or the inflammatory cardiovascularcondition in the subject. As noted above, endocrine disorders impact adiverse array of organs, and thus, the symptoms of the disorders varydepending on the tissue affected. By way of example, symptoms of theinflammatory cardiovascular condition that may be improved by decreasinginflammation or the inflammatory response, locally and/or systemically,include, but are not limited to: chest pain, irregular heart rhythm,angina, shortness of breath, dizziness, decreased activity level, andfatigue. Improvement in patients (e.g., decrease in symptoms) may bemeasured directly by assessing inflammation or by evaluating cytokineexpression in patients. Improvement can also be assessed by evaluatingimprovement in any of the foregoing symptoms, evaluating patientself-reporting of quality of life and symptom reduction, and evaluatingimprovement in activity levels.

Inflammatory conditions of the kidney include, but are not limited to,glomerulonephritis, interstitial nephritis, lupus nephritis, nephritissecondary to Wegener's disease, acute renal failure secondary to acutenephritis, Goodpasture's syndrome, post-obstructive syndrome and tubularischemia. In certain embodiments, the disclosure provides methods oftreating an inflammatory kidney condition in a patient in need thereofby administering an effective amount a compound of the disclosure. Incertain embodiments, treating an inflammatory kidney condition comprisesdecreasing inflammation in the kidney in the patient, decreasingmisregulation of inflammatory cytokines in the patient, decreasingcirculating levels of one or more cytokines, for example, IL-4, IL-10and/or IL-12, in plasma of the patient, and/or decreasing one or moresymptoms of the inflammatory kidney condition in the subject. By way ofexample, symptoms of the inflammatory kidney condition that may beimproved by decreasing inflammation or the inflammatory response,locally and/or systemically, include, but are not limited to: increasedor decreased frequency of urination, difficulty urinating, abnormallevels of protein in urine, misregulation of salt levels, blood inurine, kidney failure, and reliance on dialysis (treatment is used todecrease or eliminate reliance on dialysis). Improvement in patients(e.g., decrease in symptoms) may be measured directly by assessinginflammation or by evaluating cytokine expression in patients.Improvement can also be assessed by evaluating improvement in any of theforegoing symptoms, evaluating patient self-reporting of quality of lifeand symptom reduction, or evaluating decreased reliance on dialysis (orincreasing the period of time between diagnosis and onset of the timewhen the patient requires dialysis). Improvement can also be assessed byan increase in the period of time between diagnosis and progressing toend stage renal disease (ESRD) and/or delay or elimination of the needfor a kidney transplant. In certain embodiments, the inflammatorycondition of the kidney is an autoimmune condition, and the disclosureprovides for methods of treating such a condition.

Inflammatory conditions of the liver include, but are not limited to,hepatitis (arising from viral infection, autoimmune responses, drugtreatments, toxins, environmental agents, or as a secondary consequenceof a primary disorder), obesity, biliary atresia, primary biliarycirrhosis and primary sclerosing cholangitis. Inflammatory diseases ofthe adipose tissues include, but are not limited to, obesity. In certainembodiments, the disclosure provides methods of treating an inflammatoryliver condition in a patient in need thereof by administering aneffective amount a compound of the disclosure. In certain embodiments,treating an inflammatory liver condition comprises decreasinginflammation in the liver in the patient, decreasing misregulation ofinflammatory cytokines in the patient, decreasing circulating levels ofone or more cytokines, for example, IL-4, IL-10 and/or IL-12, in plasmaof the patient, and/or decreasing one or more symptoms of theinflammatory liver condition in the subject. By way of example, symptomsof the inflammatory liver condition that may be improved by decreasinginflammation or the inflammatory response, locally and/or systemically,include, but are not limited to: jaundice, abdominal swelling, darkurine, pale stool, bloody stool, fatigue, nausea, and loss of appetite.Improvement in patients (e.g., decrease in symptoms) may be measureddirectly by assessing inflammation or by evaluating cytokine expressionin patients. Improvement can also be assessed by evaluating improvementin any of the foregoing symptoms, evaluating patient self-reporting ofquality of life and symptom reduction. Improvement can also be assessedby a delay or elimination of the need for a liver transplant.

Inflammatory conditions of the central nervous system include, but arenot limited to, multiple sclerosis and neurodegenerative diseases suchas Alzheimer's disease, Parkinson's disease or dementia associated withHIV infection. In certain embodiments, the disclosure provides methodsof treating an inflammatory condition in a subject in need thereof, withthe proviso that the subject does not have and/or is not being treatedfor Alzheimer's disease or Parkinson's disease. In certain embodiments,the disclosure provides methods of treating an inflammatory condition ina subject in need thereof, with the proviso that the subject does nothave and/or is not being treated for an inflammatory condition of thecentral nervous system and/or a neuronal or neurodegenerative conditioncharacterized by an inflammatory component. In certain embodiments, theinflammatory condition to be treated by the methods of the disclosure isnot an inflammatory condition of the central nervous system. In certainembodiments, the inflammatory condition to be treated by the methods ofthe disclosure is not an inflammatory condition of the peripheralnervous system.

In certain embodiments, the inflammatory condition is an autoimmunedisease. Exemplary autoimmune diseases include, but are not limited to,rheumatoid arthritis, psoriasis (including plaque psoriasis), psoriaticarthritis, ankylosing spondylitis, ulcerative colitis, multiplesclerosis, lupus, alopecia, autoimmune pancreatitis, Celiac disease,Behcet's disease, Cushing syndrome, and Grave's disease. In certainembodiments, the disclosure provides methods of treating an autoimmunedisease in a patient in need thereof by administering an effectiveamount a compound, salt and/or prodrug of the disclosure.

In certain embodiments, the inflammatory condition is a rheumatoiddisorder. Exemplary rheumatoid disorders include, but are not limitedto, rheumatoid arthritis, juvenile arthritis, bursitis, spondylitis,gout, scleroderma, Still's disease, and vasculitis. We note that certaincategories of conditions overlap. For example, rheumatoid arthritis isan inflammatory rheumatoid disorder, an inflammatory joint disorder, andan autoimmune disorder. In certain embodiments, the disclosure providesmethods of treating a rheumatoid disorder in a patient in need thereofby administering an effective amount a compound, salt or prodrug of thedisclosure.

Other inflammatory conditions include periodontal disease, tissuenecrosis in chronic inflammation, endotoxin shock, smooth muscleproliferation disorders, tissue damage following ischemia reperfusioninjury, and tissue rejection following transplant surgery.

In certain embodiments, the compounds and/or compositions of thedisclosure are not for use in the treatment of Alzheimer's disease orParkinson's disease. In certain embodiments, the compounds and/orcompositions of the disclosure are not for use in the treatment of apatient who has been diagnosed with or is suspected of havingAlzheimer's disease or Parkinson's disease. In certain embodiments, thecompounds and/or compositions of the disclosure are not for use in thetreatment of a neural or neurodegenerative disease or disorder. Incertain embodiments, the compounds and/or compositions of the disclosureare not for use in the treatment of inflammatory pain. In certainembodiments, the compounds and/or compositions of the disclosure are notfor use in the treatment of pain. In certain embodiments of any of theforegoing, the term “are not for use in the treatment of” means that acompound is not being used to treat the condition and/or is not beingused with the purpose of treating the condition. In other words, incertain embodiments, the inflammatory condition being treated is notAlzheimer's disease or is not Parkinson's disease, or is not aneurodegenerative disease (in other words, is a non-neurodegenerative,inflammatory condition). Similarly, in certain embodiments, the subjectin need of treatment for an inflammatory condition, including any of theinflammatory disorders set forth herein, is not a subject diagnosed withor suspected of having Alzheimer's disease and/or Parkinson's disease.In certain embodiments, the subject in need of treatment for aninflammatory condition, including any of the inflammatory disorders setforth herein, is not a subject being treated for a neurologicalcondition or a neurodegenerative condition.

The present disclosure further provides a method of treating orpreventing inflammation associated with post-surgical wound healing in apatient.

It should be noted that the inflammatory conditions and categories ofconditions cited above are meant to be exemplary rather than exhaustive.Those skilled in the art would recognize that additional inflammatorydiseases (e.g., systemic or local immune imbalance or dysfunction due toan injury, infection, insult, inherited disorder, or an environmentalintoxicant or perturbant to the subject's physiology) may be treated bythe methods of the current disclosure.

Inflammatory conditions can be categorized by the primary tissueaffected. Illustrative examples of inflammatory conditions socategorized are provided above. The disclosure contemplates treating anysuch categories of inflammatory conditions by administering an effectiveamount of a compound, salt and/or prodrug of the disclosure to a patientin need thereof. Moreover, inflammatory conditions can be furthercategorized based on the mechanism of action underlying the condition.For example, inflammatory conditions may be categorized as autoimmune,as chronic versus acute, based on the portion of the immune system thatis hyperactivated or upregulated in the condition, or based on thecytokines or category of cytokines misregulated in the condition. Incertain embodiments, the inflammatory condition is an allergic reactionor other inflammatory response mediated by IgE antibodies. In certainembodiments, the inflammatory condition is mediated by misregulation ofinflammatory cytokines, such as interleukins (ILs) or tumor necrosisfactor alpha (TNF).

Inflammatory conditions suitable for treatment with a compound, salt orprodrug of the disclosure may also be categorized based on the one ormore cytokines that are elevated in patients (for example, in a tissueor body fluid (e.g., blood, serum or plasma) of the patient) having theconditions and/or that mediate, in whole or in part, the symptoms of thecondition. In certain embodiments, inflammatory conditions suitable fortreatment are conditions characterized, in whole or in part, by elevatedlevels (e.g., elevated levels in plasma and/or in a tissue in whichsymptoms are present) of one or more of the following cytokines: IL-4,IL-10, and/or IL-12. It should be noted that additional cytokines mayalso be elevated. However, in certain embodiments, the inflammatorycondition is characterized by elevated concentrations, such as elevatedin plasma concentrations, of at least IL-4, IL-10, and/or IL-12.Exemplary conditions that may, in certain embodiments, be characterizedby elevated levels of IL-4, IL-10 and/or IL-12 include, but are notlimited to, rheumatoid arthritis, psoriasis (including plaquepsoriasis), psoriatic arthritis, atherosclerosis, Crohn's disease,irritable bowel syndrome, ulcerative colitis, multiple sclerosis, jointautoimmune inflammation, and immune-mediated inflammatory disorders. Thedisclosure contemplates methods in which a subject in need of treatmentfor any of the foregoing conditions or any condition characterized byelevated levels of IL-4, IL-10, and/or IL-12 may be treated byadministering an effective amount of a compound of the disclosure (e.g.,a compound, salt or prodrug). In certain embodiments, the conditionbeing treated is not Alzheimer's disease and/or the subject in needthereof does not have and/or is not being treated for and/or has notbeen diagnosed with and/or is not suspected of having Alzheimer'sdisease. In certain embodiments, the condition being treated is notParkinson's disease and/or the subject in need thereof is not beingtreated for and/or has not been diagnosed with and/or is not suspectedof having Parkinson's disease. In certain embodiments, the condition ischaracterized by elevated levels of, at least, IL-12, and the disclosureprovides methods for reducing levels of IL-12, such as in the plasma, ofpatients having any of the foregoing conditions or another conditionmediated, in whole or in part, by IL-12 misregulation. Throughout thedisclosure, a reference to an increased (elevated) level orconcentration of one or more cytokines, for example, IL-12, in a subject(for example, in a tissue or body fluid sample of the subject) with aparticular condition, for example, an inflammatory condition, refers toan increased (elevated) level or concentration of the cytokine in asubject with the condition relative to a subject without the condition.

In certain embodiments, a compound, salt, or prodrug of the disclosureis administered to decrease levels of one or more cytokines in a subjectin need thereof (e.g., a subject with an inflammatory condition). Incertain embodiments, levels of cytokine are decreased in the plasma ofthe treated subject. Exemplary cytokines that may be decreased, such asdecreased in a subject in need thereof, include, but are not limited to,IL-15, IL-1b, IL-2, IL-7, IL-9, IL-10, IL-17, MIG, and MIP1a. Furtherexemplary cytokines that may be decreased, such as decreased in theplasma of treated subjects, include, but are not limited to, IL-3, IL-4,IL-10, IL-12, IFN-r, IL-5, IL-6, IL-13, and MIP1b. In certainembodiments, at least one, at least two, at least three, at least four,at least five, at least six, at least seven, at least eight, at leastnine cytokines are decreased following treatment. In certainembodiments, at least IL-4, IL-10, and/or IL-12 are reduced in treatedsubjects, such as in plasma of treated subjects. In other words, incertain embodiments, the disclosure provides a method for reducing thelevel of one or more of IL-4, IL-10, and/or IL-12 in a subject in needthereof, such as reducing cytokine levels in plasma of the subject. Incertain embodiments, at least IL-4, IL-10, and IL-12 are reduced intreated subjects, and the disclosure provides a method for reducinglevels of IL-4, IL-10, and IL-12 in a subject in need thereof. Incertain embodiments, at least IL-12 is reduced in treated subjects. Inany of the foregoing, the disclosure contemplates that one or more(e.g., 1, 2, 3, 4, 5, 6, 7, 8,9, 10, or more than 10) additionalcytokines may also be reduced following treatment. The disclosurecontemplates that, in certain embodiments, administering a compound,salt, or prodrug of the disclosure may result in a statisticallysignificant decrease in levels of a particular cytokine or of one ormore cytokines in plasma in the subject, relative to the levels prior toone or more treatments. However, the disclosure also contemplates thatthe levels of such cytokines or of other cytokines, though reducedfollowing one or more treatments, may be reduced to a lesser degree(e.g., the average level may decrease even if the total change is notstatistically significant). As is common with the administration ofcompounds, the intended affect (e.g., reduction in plasma levels ofcertain cytokines) may require multiple treatments over some period oftime. Thus, the disclosure contemplates that a reduction in cytokinelevels in plasma, or any other affect, may be observable after a singletreatment or after multiple treatments.

Without being bound by theory, the reduction in circulating cytokines inthe plasma may be mediated by actions on macrophages and monocytes,thereby indicating applicability of treatment with a compound, salt, orprodrug of the disclosure to numerous inflammatory conditions. Incertain embodiments, the inflammatory condition is mediated, in whole orin part, by elevated interleukins.

Without being bound by theory, generally suppressing the immune systemmay result in unwanted side effects. Thus, in certain embodiments,administering a compound, salt or prodrug of the disclosure to a patientto treat an inflammatory condition reduces levels of one or morecytokines, but does not generally decrease levels of all cytokines.Thus, in certain embodiments, administering a compound, salt or prodrugof the disclosure does not cause general immunosuppression. For example,in certain embodiments, although the levels of one or more cytokines aredecreased, the levels of one or more of the following are unchanged,substantially unchanged, or even slightly increased followingadministration of a compound, salt or prodrug of the disclosure: M-CSF,GM-CSF, G-CSF, MCP-1, IP-10, MIG, eotaxin, MIP-2, or LIX. In certainembodiments, administering a compound, salt or prodrug of the disclosureto a subject does not result in a statistically significant increase inthe risk of opportunistic infections versus that for subjects having thesame condition but not so treated. In certain embodiments, administeringa compound, salt or prodrug of the disclosure to a subject does notresult in neutropenia.

In certain embodiments, the condition being treated comprises rheumatoidarthritis. In certain embodiments, the patient has elevated levels ofIL-12, such as elevated levels in plasma and/or in synovial tissue.Rheumatoid arthritis is an autoimmune disease and is a chronic, systemicinflammatory disorder. Rheumatoid arthritis primarily affects thejoints, particularly the synovial joints, but it may also affect manyother tissues and organs including the lungs, pericardium, and sclera.The condition can be disabling and painful, and patients whose diseaseis not adequately managed may have significant loss of mobility andsubstantial impairments in daily functioning. Numerous animal models ofrheumatoid arthritis exist and may be used, for example, to optimizetreatment regimens. These models include the collagen-induced arthritismodel, the collagen-antibody-induced arthritis model, thezymosan-induced arthritis model, and the methylated BSA model. Inaddition, genetically manipulated transgenic mouse lines exist andprovide suitable models. For a review of numerous models see Asquith etal., 2009, European Journal of Immunology 39(8): 2040-4.

In certain embodiments, the condition being treated comprises psoriasis,such as plaque. In certain embodiments, the condition being treatedcomprises psoriatic arthritis. There are five types of psoriasis:plaque, guttate, inverse, pustular, and erythrodermic. The most commonform, plaque psoriasis, is commonly seen as red and white hues of scalypatches appearing on the epidermis. In certain embodiments, thecondition being treated herein is plaque psoriasis. Psoriasis can alsocause inflammation of the joints, which is known as psoriatic arthritis.Approximately 10-30% of patients with psoriasis also have psoriaticarthritis. In certain embodiments, the disclosure provides methods fortreating psoriatic arthritis. In certain embodiments the patient in needof treatment for psoriasis, such as plaque psoriasis, or for psoriaticarthritis has elevated levels of IL-12. Animal models of psoriasis areavailable and may be used, for example, to optimize treatment regimens.See, for example, Conrad, 2006, Current Rheumatology Report 8(5):342-347.

In certain embodiments, the condition being treated comprisesatherosclerosis. Atherosclerosis is a condition in which an artery wallthickens as a result of the accumulation of fatty materials such ascholesterol and triglyceride. It affects arterial blood vessels andinvolves a chronic inflammatory response, such as in the walls ofarteries. Animal models are available and may be used, for example, tooptimize treatment regimens. See, for example, Getz, 2012, ArteriosclerThromb Vasc Biol. 32(5): 1104-15.

In certain embodiments, the condition being treated is inflammatorybowel disease, such as Crohn's disease or ulcerative colitis. Crohn'sdisease is a type of inflammatory bowel disease that may affect any partof the gastrointestinal tract (e.g., mouth to anus), leading to diverseGI symptoms. Approximately 50% of cases affect both ileum and the largeintestines. The primary symptoms include abdominal pain, diarrhea,vomitting, and/or weight loss. In addition, patients may experiencesymptoms and complications in other tissues and organs, such as anemia,skin rash, arthritis, inflammation of the eye, and fatigue. In somecases, uncontrolled disease may lead to obstruction, fistula, orabscess. Ulcerative colitis affects the colon and is characterized byulcers or open sores. The main symptom of active disease includesconstant diarrhea mixed with blood and/or mucus. The frequency andseverity of the diarrhea varies with the severity of the disease, andthe GI-tract bleeding may lead to anemia. Like with Crohn's disease,non-GI symptoms may also be present. Severe ulcerative colitis can leadto perforation and may be fatal. Numerous animal models to, for example,study inflammatory bowel disease and/or optimize treatment areavailable. See, for example, Mizoguchi, 2012, Prog Mol Biol Transl Sci.105: 263-320.

In certain embodiments, the condition being treated comprises irritablebowel syndrome. Irritable bowel syndrome generally involves asensitization of the nerves responsible for peristalsis. As a result,the muscles controlled by these nerves spasm in response to mildstimuli, such as certain foods or stress. Symptoms include pain,diarrhea, and/or constipation.

In certain embodiments, the condition being treated, such as theinflammatory condition being treated, is endometriosis. For example, asubject, specifically a female subject, having or suspected of havingendometriosis is treated with a compound, salt or prodrug of thedisclosure. Endometriosis is a gynecological condition in which cellsfrom the lining of the uterus (endometrium) appear and flourish outsidethe uterine cavity, most commonly on the membrane which lines theabdominal cavity, the peritoneum. Symptoms of endometriosis are pain,particularly pelvic pain, and infertility. The pain often is worse withthe menstrual cycle. Endometriosis is typically seen during thereproductive years, and has been estimated to occur in roughly 6-10% ofwomen. Current treatments are primarily pain management, hormonetreatment, and surgery.

In addition to infertility, the other primary symptom of endometriosisis recurring pelvic pain. The pain can range from mild to severecramping or stabbing pain that occurs on both sides of the pelvis, inthe lower back and rectal area, and even down the legs. The amount ofpain a woman feels correlates poorly with the extent or stage (1 through4) of endometriosis, with some women having little or no pain despitehaving extensive endometriosis or endometriosis with scarring, whileother women may have severe pain even though they have only a few smallareas of endometriosis. Thus, pain is a poor indicator of the extent ofthe condition. Symptoms of the pain include: dysmenorrhea (painfulcramps during menses); chronic pelvic pain; dyspareunia (painfulintercourse); and dysuria (frequent and sometimes painful urination).

Endometriosis lesions may bleed and swell. This can trigger both aninflammatory response and pain. However, the pathophysiology of thecondition is multifactorial and aspects of that pathophysiology broadlyinclude underlying predisposing factors, inflammation, metabolicchanges, formation of ectopic endometrium, and generation of pain.Adhesions may form, thereby causing both pain and additionalcomplications such as organ dislocation. Recently, it has been shownthat there are elevated levels of IL-10, IL-12(p70), MIP1a, MIP1b, andTNFα in the supernatant of peritoneal macrophages from subjects withendometriosis. Additionally, subjects having endometriosis have beenreported to have elevated IL-10 serum levels. These findings underscorethe inflammatory component to endometriosis.

In certain embodiments, a compound, salt or prodrug of the disclosure isused to treat endometriosis, such as to reduce inflammation associatedwith endometriosis. In certain embodiments, a compound, salt or prodrugof the disclosure is used to decrease elevated IL-10, IL-12(p70), MIP1a,MIP1b, and/or TNFα levels in subjects having endometriosis, such as toreduce concentration of pro-inflammatory cytokines in the peritonealcavity and/or in the plasma, such as in circulating plasma.

In certain embodiments, the disclosure provides a method of decreasingconcentration of a pro-inflammatory cytokine (e.g., decreasing elevatedconcentrations; decreasing elevated levels), such as decreasing levelsin the plasma, by administering a compound, salt or prodrug of thedisclosure. In certain embodiments, the pro-inflammatory cytokine isselected from one or more of: IL-4, IL-12(p70), MIPa, MIPb, TNFα, IL-7,IL-13, IL-17, or IL-10. In certain embodiments, the method is performedin vitro or in vivo. In certain embodiments, the method is performed ina subject (e.g., a human or non-human animal) having an inflammatorycondition or an immune mediated disorder, such as any one or more of thediseases and conditions described herein.

In certain embodiments, a compound, salt, and prodrug of the disclosureis administered topically, for example, to decrease inflammation in aninflammatory skin disorder. In certain embodiments, a compound, salt,and/or prodrug of the disclosure is administered locally, for example,injected into the space around an inflamed joint in a subject withrheumatoid arthritis. In certain embodiments, a compound, salt, andprodrug of the disclosure is administered systemically, such as orallyor intravenously. These are merely exemplary. The appropriate route ofadministration may be selected based on the particular indication beingtreated and the patient's condition, and numerous exemplary routes ofadministration are described herein and known in the art. In certainembodiments, a compound, salt or prodrug of the disclosure isadministered orally (e.g., is orally bioavailable).

In certain embodiments, any of the methods described herein compriseproviding a composition (e.g., a pharmaceutical composition) comprisinga compound or salt of the disclosure or providing a compound of thedisclosure, which composition is formulated with an acceptable carrierand/or excipient, and delivering or otherwise administering to a subjector patient in need thereof that composition or compound. In certainembodiments, the composition is for oral delivery to a subject orpatient, and administering to a subject or patient in need thereofcomprises orally administering that composition to the subject orpatient.

The disclosure contemplates methods of treating any one or more of theforegoing diseases or conditions (including categories of diseases orconditions) using a compound, salt or prodrug of the disclosure.Similarly, the disclosure contemplates methods of treating any one ormore of the foregoing diseases or conditions (including categories ofdiseases or conditions) using a composition, such as a pharmaceuticalcomposition of the disclosure (e.g., a pharmaceutical compositioncomprising a compound, salt or prodrug of the disclosure). Thedisclosure contemplates methods of treating any one or more of theforegoing diseases or conditions (e.g., treating a subject or patient inneed of treatment for any one or more of the foregoing diseases ofconditions) using any of the compounds or compositions (e.g.,pharmaceutical compositions) of the disclosure. In certain embodiments,the disclosure contemplates methods of treating a subject or patient inneed of treatment for any one or more of the foregoing diseases andconditions (e.g., a patient having or suspected of having a particulardisease or condition), which subject or patient does not have and/or hasnot been diagnosed with and/or is not suspected of having another one ormore of the foregoing diseases and conditions.

E. Compositions and Modes of Administration

It will be appreciated that compounds and agents used in thecompositions and methods of the present disclosure preferably shouldreadily penetrate the blood-brain barrier when peripherallyadministered. Compounds which cannot penetrate the blood-brain barrier,however, can still be effectively administered directly into the centralnervous system, e.g., by an intraventricular route.

In some embodiments of this disclosure, the compound of the presentdisclosure is formulated with a pharmaceutically acceptable carrier. Inother embodiments, no carrier is used. For example, the compound asdescribed herein can be administered alone or as a component of apharmaceutical formulation (therapeutic composition; pharmaceuticalcomposition). The compound may be formulated for administration in anyconvenient way for use in human medicine. Any compound of the disclosureor salt or prodrug thereof can be provided as a composition, such as apharmaceutical composition, such as a composition having any of thefeatures described herein. Any such compound of the disclosure orcomposition of the disclosure may be used in any of the in vitro or invivo methods described herein.

Pharmaceutically acceptable carriers that may be used in thesecompositions include, but are not limited to, ion exchangers, alumina,aluminum stearate, lecithin, serum proteins, such as human serumalbumin, buffer substances such as phosphates, glycine, sorbic acid,potassium sorbate, partial glyceride mixtures of saturated vegetablefatty acids, water, salts or electrolytes, such as protamine sulfate,disodium hydrogen phosphate, potassium hydrogen phosphate, sodiumchloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, cellulose-based substances, polyethylene glycol, sodiumcarboxymethylcellulose, polyacrylates, waxes,polyethylene-polyoxypropylene-block polymers, polyethylene glycol andwool fat.

In some embodiments, the therapeutic methods of the disclosure includeadministering the composition of a compound topically, systemically, orlocally. For example, therapeutic compositions of compounds of thedisclosure may be formulated for administration by, for example,injection (e.g., intravenously, subcutaneously, or intramuscularly),inhalation or insufflation (either through the mouth or the nose) ororal, buccal, sublingual, transdermal, nasal, or parenteraladministration. The compositions of compounds described herein may beformulated as part of an implant or device, or formulated for slow orextended release. When administered parenterally, the therapeuticcomposition of compounds for use in this disclosure is preferably in apyrogen-free, physiologically acceptable form. Techniques andformulations generally may be found in Remington's PharmaceuticalSciences, Meade Publishing Co., Easton, Pa.

In certain embodiments, pharmaceutical compositions suitable forparenteral administration may comprise the compound of the presentdisclosure in combination with one or more pharmaceutically acceptablesterile isotonic aqueous or non-aqueous solutions, dispersions,suspensions or emulsions, or sterile powders which may be reconstitutedinto sterile injectable solutions or dispersions just prior to use,which may contain antioxidants, buffers, bacteriostats, solutes whichrender the formulation isotonic with the blood of the intended recipientor suspending or thickening agents. Examples of suitable aqueous andnon-aqueous carriers which may be employed in the pharmaceuticalcompositions of the disclosure include water, ethanol, polyols (such asglycerol, propylene glycol, polyethylene glycol, and the like), andsuitable mixtures thereof, vegetable oils, such as olive oil, andinjectable organic esters, such as ethyl oleate. Proper fluidity can bemaintained, for example, by the use of coating materials, such aslecithin, by the maintenance of the required particle size in the caseof dispersions, and by the use of surfactants.

A composition comprising a compound of the present disclosure may alsocontain adjuvants, such as preservatives, wetting agents, emulsifyingagents and dispersing agents. Prevention of the action of microorganismsmay be ensured by the inclusion of various antibacterial and antifungalagents, for example, paraben, chlorobutanol, phenol sorbic acid, and thelike. It may also be desirable to include isotonic agents, such assugars, sodium chloride, and the like into the compositions. Inaddition, prolonged absorption of the injectable pharmaceutical form maybe brought about by the inclusion of agents which delay absorption, suchas aluminum monostearate and gelatin.

In certain embodiments of the disclosure, compositions comprising acompound of the present disclosure can be administered orally, e.g., inthe form of capsules, cachets, pills, tablets, lozenges (using aflavored basis, usually sucrose and acacia or tragacanth), powders,granules, or as a solution or a suspension in an aqueous or non-aqueousliquid, or as an oil-in-water or water-in-oil liquid emulsion, or as anelixir or syrup, or as pastilles (using an inert base, such as gelatinand glycerin, or sucrose and acacia) and the like, each containing apredetermined amount of the compound of the present disclosure as anactive ingredient. In some embodiments, compounds of the presentinvention have good oral availability, e.g., oral availability of atleast about 50%, at least about 60%, or even at least about 70%.

In solid dosage forms for oral administration (capsules, tablets, pills,dragees, powders, granules, and the like), one or more compositionscomprising the compound of the present disclosure may be mixed with oneor more pharmaceutically acceptable carriers, such as sodium citrate ordicalcium phosphate, and/or any of the following: (1) fillers orextenders, such as starches, lactose, sucrose, glucose, mannitol, and/orsilicic acid; (2) binders, such as, for example, carboxymethylcellulose,alginates, gelatin, polyvinyl pyrrolidone, sucrose, and/or acacia; (3)humectants, such as glycerol; (4) disintegrating agents, such asagar-agar, calcium carbonate, potato or tapioca starch, alginic acid,certain silicates, and sodium carbonate; (5) solution retarding agents,such as paraffin; (6) absorption accelerators, such as quaternaryammonium compounds; (7) wetting agents, such as, for example, cetylalcohol and glycerol monostearate; (8) absorbents, such as kaolin andbentonite clay; (9) lubricants, such a talc, calcium stearate, magnesiumstearate, solid polyethylene glycols, sodium lauryl sulfate, andmixtures thereof; and (10) coloring agents. In the case of capsules,tablets and pills, the pharmaceutical compositions may also comprisebuffering agents. Solid compositions of a similar type may also beemployed as fillers in soft and hard-filled gelatin capsules using suchexcipients as lactose or milk sugars, as well as high molecular weightpolyethylene glycols and the like.

Liquid dosage forms for oral administration include pharmaceuticallyacceptable emulsions, microemulsions, solutions, suspensions, syrups,and elixirs. In addition to the compound of the present disclosure, theliquid dosage forms may contain inert diluents commonly used in the art,such as water or other solvents, solubilizing agents and emulsifiers,such as ethyl alcohol (ethanol), isopropyl alcohol, ethyl carbonate,ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol,1,3-butylene glycol, oils (in particular, cottonseed, groundnut, corn,germ, olive, castor, and sesame oils), glycerol, tetrahydrofurylalcohol, polyethylene glycols and fatty acid esters of sorbitan, andmixtures thereof. Besides inert diluents, the oral compositions can alsoinclude adjuvants such as wetting agents, emulsifying and suspendingagents, sweetening, flavoring, coloring, perfuming, and preservativeagents.

Suspensions, in addition to the active compounds may contain suspendingagents such as ethoxylated isostearyl alcohols, polyoxyethylenesorbitol, and sorbitan esters, microcrystalline cellulose, aluminummetahydroxide, bentonite, agar-agar and tragacanth, and mixturesthereof.

A person of ordinary skill in the art, such as a physician, is readilyable to determine the required amount of the compound of the presentdisclosure to treat the subject using the compositions and methods ofthis disclosure. It is understood that the dosage regimen will bedetermined for an individual, taking into consideration, for example,various factors that modify the action of a compound of the presentdisclosure, the severity or stage of the disease, route ofadministration, and characteristics unique to the individual, such asage, weight, size, and extent of cognitive impairment.

It is well-known in the art that normalization to body surface area isan appropriate method for extrapolating doses between species. Tocalculate the human equivalent dose (HED) from a dosage used in thetreatment of age-dependent cognitive impairment in rats, the formula HED(mg/kg)=rat dose (mg/kg)×0.16 may be employed (see Estimating the SafeStarting Dose in Clinical Trials for Therapeutics in Adult HealthyVolunteers, December 2002, Center for Biologics Evaluation andResearch). For example, using that formula, a dosage of 10 mg/kg in ratsis equivalent to 1.6 mg/kg in humans. This conversion is based on a moregeneral formula HED=animal dose in mg/kg×(animal weight in kg/humanweight in kg)^(0.33). Similarly, to calculate the HED can be calculatedfrom a dosage used in the treatment in mouse, the formula HED(mg/kg)=mouse dose (mg/kg)×0.08 may be employed (see Estimating the SafeStarting Dose in Clinical Trials for Therapeutics in Adult HealthyVolunteers, December 2002, Center for Biologics Evaluation andResearch).

In certain embodiments of the disclosure, the dose of the compound orcomposition of the present disclosure is between 0.00001 and 100mg/kg/day (which, given a typical human subject of 70 kg, is between0.0007 and 7000 mg/day). Desired duration of administration of thecompound described herein can be determined by routine experimentationby one skilled in the art. For example, the compound of the presentdisclosure may be administered for a period of 1-4 weeks, 1-3 months,3-6 months, 6-12 months, 1-2 years, or more, up to the lifetime of thepatient. For example, daily administration of the compounds over thisperiod is contemplated.

In addition to compound of the present disclosure, the compositions andmethods of this disclosure can also include other therapeutically usefulagents. These other therapeutically useful agents may be administered ina single formulation, simultaneously or sequentially with the compoundof the present disclosure according to the methods of the disclosure.

It will be understood by one of ordinary skill in the art that thecompositions and methods described herein may be adapted and modified asis appropriate for the application being addressed and that thecompositions and methods described herein may be employed in othersuitable applications, and that such other additions and modificationswill not depart from the scope hereof. For example, the compounds of thedisclosure are also useful as agents for agonizing P₂Y₆ receptoractivity, and can be used in vitro or in vivo to study normal andabnormal P₂Y₆ receptor function. In certain embodiments, the compoundsof the disclosure are used, directly or indirectly, to agonize P₂Y₆receptor activity, and may be used in any of the in vitro and/or in vivomethods disclosed herein. In certain embodiments, compounds disclosedherein are themselves P₂Y₆ receptor-modulating compounds, and thedisclosure encompasses these compounds as well as their salts and/orprodrugs as agonists of the disclosure. Other compounds, salts, andprodrugs described herein are not active themselves, but are convertedin vivo to compounds that are active P₂Y₆ receptor-modulating compounds.The disclosure contemplates that all such compounds, salts, or prodrugsof the disclosure, whether active themselves or are converted intoactive compounds in vivo, may be used to treat any of conditionsdescribed herein.

This disclosure will be better understood from the Experimental Detailswhich follow. However, one skilled in the art will readily appreciatethat the specific methods and results discussed are merely illustrativeof the disclosure as described more fully in the embodiments whichfollow thereafter.

EXAMPLES Example 1 Preparation of Compound 6

Scheme 2 below provides a general synthetic route for the preparation ofcompound 6.

Step 1: Synthesis of Compound 47

To a solution of compound 46 (3.0 g, 8.11 mmol) in DMF (90 mL) was addedY01 (3.0 g, 16.22 mmol) and K₂CO₃ (4.47 g, 16.22 mmol), the resultingmixture was stirred at 70° C. for 1 h. After cooling down, the mixturewas diluted with 250 mL water, extracted with ethyl acetate (EA) (250mL×3), the organic layer was dried over anhydrous Na₂SO₄, concentratedto give a crude product. The crude product was purified on column(eluted with PE/EA=3:1) to give 3.61 g 47 as a colorless oil, yield:94%. ¹H NMR (300 MHz, CDCl₃) δ 7.36 (d, J=8.1 Hz, 1H), 7.32-7.27 (m,4H), 7.25-7.18 (m, 1H), 5.98 (d, J=4.0 Hz, 1H), 5.81 (d, J=8.1 Hz, 1H),5.34 (d, J=2.4 Hz, 2H), 4.35 (s, 3H), 4.13 (m, 2H), 3.01-2.84 (m, 2H),2.14 (dd, J=12.1, 4.2 Hz, 9H), 1.26 (t, J=7.1 Hz, 1H).

Step 2: Synthesis of Compound 6

3.61 g 47 was dissolved in 150 mL 5N NH₃/methanol then stirred at roomtemperature for 12 hrs. After the reaction was finished, methanol wasremoved under vacuum to give the crude product. The crude product wasrecrystallized from EA to give 1.94 g 6 as a white solid, yield: 73%. ¹HNMR (300 MHz, DMSO) δ 7.95 (d, J=8.1 Hz, 1H), 7.37-7.11 (m, 5H), 5.77(m, 2H), 5.42 (d, J=5.4 Hz, 1H), 5.12 (m, 1H), 4.06-3.88 (m, 4H), 3.84(m, 1H), 3.64 (m, 1H), 3.53 (m, 1H), 2.80 (t, J=9.0 Hz, 2H).

Example 2 Preparation of Compound 3

Scheme 3 below provides a general synthetic route for the preparation ofcompound 3.

Step 1: Synthesis of Compound 49

Compound 49 was prepared from compound 46 according to the sameprocedure as described in step 1 of Example 1. 2.98 g compound 49 wasobtained from 3.0 g compound 46, yield: 79.7%. ¹H NMR (300 MHz, CDCl₃) δ8.47 (d, J=4.7 Hz, 1H), 7.58 (m, 1H), 7.43 (d, J=8.2 Hz, 1H), 7.19-7.07(m, 2H), 6.01 (d, J=4.8 Hz, 1H), 5.85 (d, J=8.2 Hz, 1H), 5.36-5.26 (m,2H), 5.20 (s, 2H), 4.31 (s, 3H), 2.05 (t, J=10.5 Hz, 9H).

Step 2: Synthesis of Compound 50

Compound 3 was prepared from compound 49 according to the same procedureas described in step 2 of Example 1. 1.79 g compound 3 was obtained from2.98 g compound 49, yield: 82.7%. ¹H NMR (300 MHz, DMSO) δ 8.42 (d,J=3.5 Hz, 1H), 8.03 (d, J=8.1 Hz, 1H), 7.76-7.67 (m, 1H), 7.26-7.18 (m,2H), 5.81 (dd, J=14.9, 6.5 Hz, 2H), 5.44 (d, J=5.7 Hz, 1H), 5.23-5.01(m, 4H), 4.03 (m, 1H), 3.96 (m, 1H), 3.84 (m, 1H), 3.70-3.59 (m, 1H),3.53 (m, 1H).

Example 3 Preparation of Compound 4

Scheme 4 below provides a general synthetic route for the preparation ofcompound 4.

Step 1: Synthesis of Compound 51

To a solution of compound 46 (1.061 g, 1.87 mmol), Y03 (930 mg, 5.73mmol) and PPh₃(1.501 g, 5.73 mmol) in 25 mL THF was added dropwise asolution of DIAD (1.159 g, 5.73 mmol) in 5 mL THF over 30 min, theresulting mixture was stirred at 50° C. for 3 h. After the reaction wasfinished, THF was removed to give the crude product. The crude productwas purified on column (eluted with EA) to give 1.37 g compound 51 as anoil, yield: 88.8%.

Step 2: Synthesis of Compound 4

Compound 4 was prepared from compound 51 according to the same procedureas described in step 2 of Example 1. 0.8 g compound 4 was obtained from1.37 g compound 51, yield: 77.4%. ¹H NMR (300 MHz, DMSO) δ 7.99 (d,J=8.1 Hz, 1H), 7.73 (d, J=8.2 Hz, 1H), 7.55 (d, J=8.5 Hz, 1H), 7.36 (t,J=7.7 Hz, 1H), 7.09 (t, J=7.5 Hz, 1H), 5.87-5.76 (m, 2H), 5.38 (d, J=5.7Hz, 1H), 5.30 (d, J=4.0 Hz, 2H), 5.12-5.06 (m, 1H), 4.01 (t, J=5.2 Hz,1H), 3.94 (s, 4H), 3.84 (d, J=3.6 Hz, 1H), 3.68-3.46 (m, 2H).

Example 4 Preparation of Compound 1

Scheme 5 below provides a general synthetic route for the preparation ofcompound 1.

Step 1: Synthesis of Compound 53

Compound 53 was prepared from compound 46 according to the sameprocedure as described in step 1 of Example 1. A crude product ofcompound 53 was obtained from 1.14 g compound 46. The crude product wasused in the next step directly without further purification.

Step 2: Synthesis of Compound 1

Compound 1 was prepared from compound 53 according to the same procedureas described in step 2 of Example 1. 700 mg compound 1 was obtained fromthe 1.14 g compound 53, yield: 59.2%. ¹H NMR (300 MHz, DMSO) δ 7.99 (d,J=9.0 Hz, 1H), 7.46-7.52 (m, 5H), 5.72-5.82 (m, 2H), 5.07-5.10 (m, 1H),4.45-4.55 (m, 2H), 3.92-4.00 (m, 2H), 3.86 (s, 1H), 3.54-3.64 (m, 2H),3.30-3.32 (m, 1H).

Example 5 Preparation of Compound 5

Scheme 5 below provides a general synthetic route for the preparation ofcompound 5.

Step 1: Synthesis of Compound 55

Compound 55 was prepared from compound 46 according to the sameprocedure as described in step 1 of Example 1. 4.2 g compound 55 wasobtained from 3.0 g compound 46, yield: 100%. ¹H NMR (300 MHz, CDCl₃) δ7.79 (d, J=8.0 Hz, 1H), 7.53 (dd, J=3.7, 1.6 Hz, 2H), 7.45 (d, J=8.2 Hz,1H), 7.30 (m, 1H), 6.04 (d, J=4.7 Hz, 1H), 5.89 (d, J=8.2 Hz, 1H), 5.50(d, J=1.7 Hz, 2H), 5.33 (m, 2H), 4.34 (d, J=4.3 Hz, 3H), 2.10 (d, J=6.6Hz, 6H), 2.04 (s, 3H).

Step 2: Synthesis of Compound 5

Compound 5 was prepared from compound 55 according to the same procedureas described in step 2 of Example 1. 2.36 g compound 5 was obtained from4.2 g compound 55, yield: 75.6%. ¹H NMR (300 MHz, DMSO) δ 8.06 (d, J=8.2Hz, 2H), 7.86 (d, J=8.0 Hz, 2H), 7.76-7.61 (m, 4H), 7.39 (t, J=7.4 Hz,2H), 5.89 (d, J=7.9 Hz, 2H), 5.80 (m, 2H), 5.38-5.42 (m, 3H), 5.16 (m,1H), 3.85-4.04 (m, 2H), 3.50-3.66 (m, 2H).

Compounds 32-37 were prepared according to similar synthetic proceduresas those used for preparing compound 5(see Scheme 5 above). Thecharacterization of these sodium salts are summarized in Table 1 below.

TABLE 1 Characterization of compounds 32-37: Compound Characterization

Yield: 97% as a solid TLC information: (DCM/MeOH = 10/1) Material: R_(f)= 0.5 Product: R_(f) = 0.2 ¹H NMR (400 MHz, MeOD): δ 8.15 (1H, d, J = 8Hz), 7.93-7.89 (1H, m), 7.43 (1H, d, J = 8.8 Hz), 7.30- 7.26 (1H, m),7.23-7.18 (1H, m), 5.94 (1H, d, J = 4.0 Hz), 5.87 (1H, d, J = 8 Hz),5.51 (2H, s), 4.19-4.17 (1H, m), 3.88 (1H, d, J = 8.8 Hz), 3.78-3.75(1H, m), 3.33-3.15 (1H, m).

Yield: 64% as a solid TLC information: (DCM/MeOH = 10/1) Material: R_(f)= 0.5 Product: R_(f) = 0.2 ¹H NMR (400 MHz, MeOD): δ 8.15 (1H, d, J = 8Hz), 7.69 (1H, d, J = 8 Hz), 7.45-7.37 (2H, m), 5.94 (1H, d, J = 4.0Hz), 5.86 (1H, d, J = 8 Hz), 5.54 (2H, s), 4.20-4.17 (1H, m), 4.03 (1H,d, J = 4 Hz), 3.86-3.77 (1H, m), 3.33-3.15 (1H, m).

Yield: 93% as solid TLC information: (DCM/MeOH = 10/1) Material: R_(f) =0.5 Product: R_(f) = 0.2

Yield: 75% as a solid TLC information: (DCM/MeOH = 10/1) Material: R_(f)= 0.5 Product: R_(f) = 0.2 ¹H NMR (400 MHz, MeOD): δ 8.15 (1H, d, J = 8Hz), 7.64 (1H, s), 7.52-7.46 (2H, m), 5.95 (1H, d, J = 4.0 Hz), 5.87(1H, d, J = 8 Hz), 5.50 (2H, s), 4.20-4.17 (1H, m), 4.04 (1H, d, J = 4Hz), 3.86-3.77 (1H, m), 3.33-3.15 (1H, m).

Yield: 83% as a solid TLC information: (DCM/MeOH = 10/1) Material: R_(f)= 0.5 Product: R_(f) = 0.2 ¹H NMR (400 MHz, MeOD): δ 8.15 (1H, d, J = 8Hz), 7.72 (1H, d, J = 8 Hz), 7.43 (1H, s), 7.21 (1H, d, J = 8.0 Hz),5.95 (1H, d, J = 4.0 Hz), 5.87 (1H, d, J = 8.0 Hz), 5.49 (2H, s),4.19-4.17 (2H, m), 4.04 (1H, d, J = 4.0 Hz), 3.87-3.77 (1H, m),3.36-3.33 (1H, m), 2.53 (3H, s).

Yield: 74% as a solid TLC information: (DCM/MeOH = 10/1) Material: R_(f)= 0.5 Product: R_(f) = 0.2 ¹H NMR (400 MHz, MeOD): δ 8.15 (1H, d, J = 8Hz), 7.66 (1H, d, J = 8 Hz), 7.43 (1H, d, J = 7.2 Hz), 7.30-7.26 (1H,m), 5.94 (1H, d, J = 4.0 Hz), 5.87 (1H, d, J = 8.0 Hz), 5.52 (2H, s),4.20-4.16 (2H, m), 4.04 (1H, s), 3.87-3.85 (1H, m), 3.78- 3.75 (1H, m),2.55 (3H, s).

Example 6 Materials and Methods for In Vitro and In Vivo Studies

Activation of P₂Y₆ Receptor

Synthetic ligands were tested for activation of P₂Y₆ receptor bymeasuring receptor induced Ca²⁺ changes with the fluorescent Ca²⁺indicator fluo-4. 1321N1 human astrocytoma cell lines either expressingP₂Y₂, P₂Y₄ or P₂Y₆ receptors were plated into 24-well plates. Two daysafter plating, fluorometric measurements were made and responses ofcells to a serial dilution of ligands were determined. P₂Y₆ receptormediated Ca²⁺ fluorescent change was determined by normalizedaccumulation of fluorescent change of 3 timepoints after ligandadministration subtracted by value from ACSF control. Changes influorescent intensity were plotted corresponding to ligand concention inGraphPad. Dose-response curve and EC₅₀ for each ligand was estimatedusing nonlinear curve fit and Sigmoidal dose-response analysis. Thesodium salt of the diphosphate derivative of compound 5 exhibited anEC₅₀ of 12 nM. The sodium salt of the diphosphate derivative of compound5 was demonstrated to selectively activate P₂Y₆ receptors by comparingits Ca²⁺ mobilizing effects in three 1321N1 human astrocytoma cell linesexpressing P₂Y₂, P₂Y₄ or P₂Y₆ receptors. The sodium salt of thediphosphate derivative of compound 5 was only effective at elevatingCa²⁺ levels when applied to cells expressing P₂Y₆ receptors and noteffective in P₂Y₂, or P₂Y₄ receptor expressing cells. The ability of thesodium salt of the diphosphate derivative of compound 5 to elevate Ca²⁺signals in P₂Y₆ receptor expressing cells was attenuated by addition ofthe P₂Y₆ antagonist MRS2578.

PSAPP Mice

Heterozygous mutant (K670N/M671L) APP (50% C57B6, 50% SJL) transgenicmice were crossed with heterozygous mutant (A246E) PS-1 (50% C57B6, 50%SJL) transgenic mice to generate heterozygous PSAPP transgenic mice(also referred to as PS-1/APP or PSAPP+/+ mice), which refers to animalsheterozygous for the PS-1 A246E transgene and the APP K670N/M671Ltransgene. Non-transgenic control animals were littermates (alsoreferred to as PSAPP_−/− mice) generated in the breeding for PSAPPtransgenic animals. Mouse genotype was determined by Polymerase ChainReaction (PCR). Both male and female mice of 6-7 months old were usedfor the experiments below. All animal experiments were performed inaccordance with the Tufts Animal Care and Use Committee and withnational regulations and policies.

Two-Photon In Vivo Imaging Study

In this study, PSAPP mice were anesthetized using isoflurane and athin-skull preparation was used to minimize the surface damage. Amyloidplaques were visualized with methoxyX04 labeling and blood plasma waslabeled with Rhodamine dextran to facilitate re-localization of the sameimaging area. Stack images were obtained using a two-photon system(Prairie Technologies) with excitation at 850 nm. The emission wasdetected by external photomultiplier tubes (525/70; DLCP 575; 607/45nm).

Stereotaxic Injection

Animals were anesthetized and immobilized in a stereotaxic frame. Foreach injection, 1 μl of 10 mM UDP or other suitable compounds inartificial cerebrospinal fluid (ACSF) as the vehicle were injectedintraventricularly using the following coordinates: AP 0.2 mm, ML 1 mm,and DV 2.2 mm.

Histology and Immunohistochemistry

Mice were perfused transcardially with 4% paraformaldehyde and 40 μmCoronal sections were collected. Sections were sequentially incubated in0.3% H₂O₂ for 10 minutes, blocking solution for 2 hrs, blocking solutioncontaining the primary antibody (rabbit anti-beta1-42; rabbitanti-beta1-40, from Chemicon International and rat anti-CD45) for 48hours at 4° C., and blocking solution containing biotinylated antibodyor fluorescently-labeled antibody for 2 hours at room temperature.Sections were visualized in a bright field microscope or a confocalmicroscope, and the optical density was obtained using MetaMorphsoftware.

Fear Conditioning Test

On day one, animals were trained in a fear conditioning apparatus for atotal of 7 minutes with a two-pairing paradigm of cue and mild footshock (a 30-s acoustic-conditioned stimulus, 80 dB; a 2-s shockstimulus, 0.5 mA). To evaluate contextual fear learning, the animalswere returned to the training context 24 hours post-training, andfreezing behavior was scored for 5 minutes. Freezing behavior wasmonitored by MotorMonitor (Hamilton Kinder) and scored every 5 seconds.

Electrophysiology and Long-Term Potentiation (LTP) Recording

Hippocampal slices (350 μm thick) were prepared from 6-month-old PSAPPmice. Baseline responses were obtained every 10 seconds and Input-output(I/O) curves, paired-pulse modification and LTP were successivelymeasured. The stimulation intensity was set to a level that gives avalue of 30% of the maximum obtained. LTP were induced by high frequencystimulation (HFS, 100 pulses at 100 Hz, four times) or by theta-burststimulation (TBS, 10 bursts at 5 Hz, repeated 10 times in 15 sintervals).

Example 7 Dose-Dependent Activation of P₂Y₆ Receptor

Synthetic ligands were tested for activation of P₂Y₆ receptor bymeasuring receptor induced Ca²⁺ changes with the fluorescent Ca²⁺indicator fluo-4, and results are shown in FIG. 10(A)-(K). 1321N1 humanastrocytoma cell lines either expressing P₂Y₂, P₂Y₄ or P₂Y₆ receptorswere plated into 24-well plates. Two days after plating, fluorometricmeasurements were made and responses of cells to a serial dilution ofligands were determined. P₂Y₆ receptor mediated Ca²⁺ fluorescent changewas determined by normalized accumulation of fluorescent change of 3timepoints after ligand administration subtracted by value from ACSFcontrol. Changes in fluorescent intensity were plotted corresponding toligand concentration in GraphPad. Dose-response curve and EC₅₀ for eachligand was estimated using nonlinear curve fit and Sigmoidaldose-response analysis. The sodium salt of the diphosphate derivative ofcompound 5 exhibited an EC₅₀ of 12 nM. The sodium salt of thediphosphate derivative of compound 5 was demonstrated to selectivelyactivate P₂Y₆ receptors by comparing its Ca²⁺ mobilizing effects inthree 1321N1 human astrocytoma cell lines expressing P₂Y₂, P₂Y₄ or P₂Y₆receptors. The sodium salt of the diphosphate derivative of compound 5was only effective at elevating Ca²⁺ levels when applied to cellsexpressing P₂Y₆ receptors and not effective in P₂Y₂, or P₂Y₄ receptorexpressing cells. The ability of the sodium salt of the diphosphatederivative of compound 5 to elevate Ca²⁺ signals in P₂Y₆ receptorexpressing cells was attenuated by addition of the P₂Y₆ receptorantagonist MRS2578. These experiments demonstrated that the diphosphatederivative of compound 5 is a P₂Y₆ receptor agonist.

Example 8 Acute UDP Administration Reduced Plaque Burden in PSAPP Mice

To evaluate the effect of UDP on plaque burden, two-photon microscopywas used to assess the amyloid plaques in the barrel cortex in livingPSAPP mice. Amyloid plaques were stained by systemically administeredmethoxy-X04. One day prior to imaging, PSAPP mice were injected withmethoxyX04 to label the amyloid plaques. On the imaging day, tofacilitate the re-location of the same imaging area, blood plasma waslabeled with Rhodamine dextran. Images were obtained from the samestart- and end-point to ensure the same image volume.

The results were shown in a maximum intensity projection of afluorescent stack containing 45 planes. Representative images ofmethoxyX04 labeled amyloid plaques and angiopathy on days 1 are shown inFIG. 1(A)-(C). Immediately after imaging, animals were injected withACSF or UDP intracerebroventricularly (i.c.v.) and allowed to recover.On day 4, animals were subjected to a second period of imaging of thesame regions studied on day 1 and the results are shown in FIG.1(D)-(F). The similar pattern of angiopathy (shown by open arrows)indicated the same imaging area.

Overall, decreased plaque occupied-area was observed on day 4 followingadministration of UDP. In the images with higher magnification (FIGS.1(C) and (F)), the same dense core plaques (as shown by arrows) could beidentified based on its morphology and location relative to the bloodvessel. It was observed that the dense core plaques had more intensemethoxyX04 labeling, but with decreased plaque size (as shown byarrows), when compared to the size of the same plaques on day 1. Thissuggested that acute UDP treatment reduced plaques size in live animals.This effect was further evaluated by quantifying the number of plaques,plaque load, and size of cross-section of individual plaques. See FIG.2(A)-(E). Quantitative analysis showed that acute UDP treatment led to a12.6% reduction in the number of plaques (P<0.01) and a 17.2% reductionin plaque load (P<0.01) in barrel cortex as assessed by two-photonmicroscopy. Individual identified plaques that were detected on thesecond imaging session showed an 18.2% reduction (P<0.01) incross-sectional area following UDP treatment.

After repeated imaging, brains were fixed and subjected to postmortemimmunohistochemistry with amyloid beta specific antibodies β1-40 andβ1-42 to evaluate the plaque load (area occupied by immunostaining ofplaque) in cortex and hippocampus. See FIG. 3(A)-(D). UDP treatmentresulted in a 60% (p<0.05) and 62% (p<0.01) decrease in plaque load inthe cortex and hippocampus, respectively, as assessed by staining withthe β1-40 antibody. Quantification of staining with β1-42 antibodyshowed a 48% (P<0.01) and 47% (P<0.05) decrease in plaque load in thecortex and hippocampus, respectively. See FIG. 4(A)-(F). Both in vivoimaging and post hoc staining showed decrease in plaque burden in brainsof PSAPP mice, consistent with reduced plaque load in the tested animalsfollowing acute administration of UDP (e.g., a P₂Y₆ receptor agonist).

Example 9 Activation of P₂Y₆ Receptors Reduced Plaque Burden in PSAPPMice

3-phenacyl-UDP (also referred to as PSB0474) is a potent and selectiveP₂Y₆ receptor agonist (EC50=70 nM, >500-fold selective). In this study,P₂Y₆ receptor was activated in vivo using 3-phenacyl-UDP (PSB0474). Theeffect of this activation may have on plaque burden was also evaluated.

PSB0474 was systemically administered to PSAPP mice via intraperitonealinjection for 2, 4 and 6 consecutive days. In one group, prior toevaluation and following to administration for 6 consecutive days,treatment was suspended for two weeks (6+2 weeks group). Brains werethen fixed and plaque load was evaluated by immunostaining with theamyloid beta specific antibodies: β1-40 and β1-42. Representative imagesof plaque load in cortex and hippocampus from animals that receivedinjections of PSB0474 according to the foregoing injection schedules areshown in FIG. 5(A)-(D). Quantitative data showed that administration ofPSB0474 for 4 and 6 consecutive days significantly decreasedimmunoreactivity of β1-40 in both cortex and hippocampus (FIGS. 6(A) and6(B)). Whereas, when administration of PSB0474 was stopped for 2 weeksfollowing six consecutive days of treatment (denoted as the 6+2 weeksgroup), β1-40 staining rebounded; although to a level lower thanobserved in mice treated with saline as a vehicle control. FIGS. 6A and6B depict the reduction in plaque load (%) the cortex and hippocampus,respectively, in PSAPP mice after treatment with 3-phenacyl-UDP for 2,4, or 6 consecutive days, as assayed by staining with the β1-40antibody. FIGS. 6C-6F depict data obtained following administration ofdifferent dosages of PSB0474. It is important to note that a 1000×increase in dose of PSB0474 did not cause detrimental effects to theanimal, suggesting that there is a wide therapeutic window for P₂Y₆receptor agonists. However, with the higher dose of 1 mg/kg we didobserve smaller effects on the efficacy endpoint presumably because theenhanced receptor occupancy led to some desensitization/internalizationof the P₂Y₆ receptor. This result indicates that activation of P₂Y₆receptor significantly attenuated plaque load in both the cortex andhippocampus in PSAPP mice.

Example 10 Acute UDP Administration Improved Cognitive Function andHippocampal LTP in PSAPP Mice

Amyloid beta peptide has been reported to be toxic to synaptictransmission, and accumulation of amyloid protein is associated withcognitive impairment both in animal models of AD and in AD patients.Additionally, accumulation of amyloid protein is observed in otherconditions associated with cognitive impairment, such as in DownSyndrome. Therefore, we further investigated in PSAPP mice whether theobserved reduction in plaque burden would also lead to reversal incognitive and memory deficits typically observed in AD patients, such asimpaired cognition, impaired memory, and deficits in long-termpotentiation (LTP).

In this study, the fear conditioning associative learning paradigm wasused as a rapid cognition assay for PSAPP mice. This study allowed us toprobe cognitive function with a single training day followed in 24 hoursby tests for contextual and cued fear learning. Contextual fear learningis dependent upon a brain area that has been implicated as a locus forcognitive decline in AD: the hippocampus. Two pairings of CS-US for fearconditioning were followed 24 hours later by testing for contextual andcued fear learning. Previous studies have reported that PSAPP animalsappear to have a selective hippocampus-dependent impairment inassociative learning following two pairings of conditioned stimuli forfear conditioning.

In this study, it was found that PSAPP mice treated with ACSF showed lowfreezing behavior during 5 minute-testing time (FIG. 7(A)), which issimilar to the level reported in previous study (Dineley, et al. 2002).After UDP treatment, PSAPP mice exhibited increased freezing behaviorduring the first 4 minutes but not during the last minute. Analysis oftotal freezing percentage (FIGS. 7(B) and 7(C)) showed that PSAPP micetreated with acute UDP exhibited significantly higher freezing behavior(49%±5%) compared to an animal treated with ACSF (18%±3%). This datasuggested that acute UDP treatment rescued the deficit in contextualfear learning in PSAPP mice.

In the fear conditioning test mice exhibit a freezing behavior if theyhave a memory of the application of the aversive shock that wasdelivered 24 hours earlier. When placed in the appropriate environmentthe mice “freeze” and do not explore their environment as theyanticipate the delivery of an additional shock. Thus the greater percenttime that they exhibit freezing indicates a greater memory of theirprevious experience and thus improved memory. This represents a decreasein the cognitive impairment observed in the untreated mice.

Accumulated evidence has shown that amyloid peptides naturally secretedor isolated from Alzheimer's brains impair synaptic plasticity,especially hippocampal long-term potentiation (Walsh et al., 2002).Therefore, we further performed LTP recordings in PASPP mice andinvestigated whether P₂Y₆ receptor-mediated plaque clearance affectssynaptic plasticity. In this study, LTP was successfully induced in CA1area of the hippocampus in aged PSAPP mice with high-frequencystimulation (HFS, 100 pulses at 100 Hz, four times in 20 s intervals).First, it was observed that LTP at the schaffer collateral synapsewithin the CA1 region was depressed in PSAPP mice, as compared withlittermates (FIG. 8(A)). This result confirmed previous reports aboutsynaptic toxicity of Abeta. Acute UDP treatment reversed this LTPdeficit in PSAPP mice, and the LTP significantly increased compared withmice injected with ACSF (FIG. 8(B)). Analysis of the last 15 minpotentiation showed a significant increase in field excitatorypostsynaptic potential (fEPSP) in PSAPP mice treated with UDP, which iscomparable to the level in PSAPP littermates (FIG. 8(C)). These datasupports the conclusion that activation of P₂Y₆ receptor rescues the LTPdeficiency in PSAPP mice, which is consistent with improvement incognition mediated by P₂Y₆ receptor.

Example 11 Activation of P₂Y₆ Receptor with Chronic Injection of PSB0474Improved Cognitive Function of PSAPP Mice

Similar to acute UDP treatment, chronic injection of the P₂Y₆ receptoragonist 3-phenacyl-UDP (PSB0474) increased total freezing percentage incontext test in PSAPP mice (FIG. 9(A)-(C)). In this study, PSB0474 wasadministered at two different doses, both of which showed beneficialeffect in improving cognitive function in the PSAPP mice.

Example 12 Activation of P₂Y₆ Receptor with the Diphosphate Derivativeof Compound 5 Improved Cognitive Function of PSAPP Mice and ReducedPlaque Burden in PSAPP Mice

In this study, the diphosphate derivative of compound 5 was injectedintraperitoneally into 6 to 7-month-old PSAPP and WT mice daily at twodifferent doses, i.e., 1 μg/kg or 1 mg/kg of the diphosphate derivativeof compound 5 (in 1% DMSO/PBS) for 7 consecutive days. Consistent withthe results observed following acute UDP or PSB0474 treatment, treatmentwith the diphosphate derivative of compound 5 increased total freezingpercentage in the context test in PSAPP mice (FIG. 11). FIG. 11 showsfreezing behavior (freezing %) of PASPP mice in fear conditioningstudies after treatment with vehicle control or the diphosphatederivative of compound 5. FIG. 11 depicts the results of experimentsusing the contextual fear conditioning test with PSAPP mice treated withvehicle control (black bar at center of graph). These mice showedsignificantly decreased freezing percentage compared to the age-matchedwildtype animals (white bar); indicative of the memory deficits andcognitive impairment in PSAPP mice. Administration of the diphosphatederivative of compound 5 prior to testing significantly improved thefreezing behavior (hatched bar at right of graph) compared to thecontrol treatment. In fact, this behavior which is indicative ofcognitive function and memory was restored to a level equivalent to thatobserved in wildtype animals. This result is consistent with theconclusion that the diphosphate derivative of compound 5 improvedcognitive function (decreased cognitive deficits) in these mice, such asby improving memory and/or learning.

Treatment with the diphosphate derivative of compound 5 was also foundto reduce the plaque burden in cortex and hippocampus of PSAPP mice(FIG. 12 (A)-(C)). FIG. 12 shows plaque load in the cortex (Cx) andhippocampus (Hp) of the PSAPP mice after treatment with the diphosphatederivative of compound 5 or vehicle control, as assayed using theamyloid beta specific antibody β1-42. FIG. 12A depicts the substantialdecrease in Aβ plaque load (%) in the cortex following treatment withthe diphosphate derivative of compound 5, in comparison to the vehiclecontrol. FIG. 12B depicts the substantial decrease in Aβ plaque load (%)in the hippocampus following treatment with the diphosphate derivativeof compound 5, in comparison to the vehicle control. FIG. 12C showspostmortem immunohistochemistry analysis of the plaque load in cortexand hippocampus of PSAPP mice after treatment with the diphosphatederivative of compound 5 or vehicle control. Amyloid beta specificantibody β1-42 was used in the analysis.

To generate these graphs showing plaque load, mice were euthanized,brain sections cut and antibodies directed against Aβ 42 were used todisclose Aβ plaques. Images were acquired digitally and an algorithm wasapplied to threshold the image so that plaques were isolated from thebackground. The algorithm then calculated the percent area of the fieldof view occupied by the plaques.

Example 13 Nucleoside Compound 5 Reduces Plaque Burden in PS1/APP Mice

In this study, the effect of systemic administration of nucleosidecompound 5 on reducing plaque burden in PS1/APP mice was examined.

Animal Groups:

I.P. Injection Animal Group Compound Dose time frame 1) PS1/APP+/+ GroupA 10 μg/kg Once per day for (26 weeks, n = 7) compound 5 7 days 2)PS1/APP+/+ Group B vehicle Once per day for (26 weeks, n = 8) 7 days

Brain samples were collected for Abeta analysis with immunostaining.Brains samples were collected after transcardial perfusion withphosphate buffer and 4% paraformaldehyde. Floating brain sections (40micron) were collected using a microtome. Immunostaining for Aβ40 andAβ42 were performed according to previously established protocol. Imageswith DAB staining were collected with brightfield microscopy andanalyzed with ImageJ Macro. All data analysis and statistics wereperformed in GraphPad Prism 6.0.

The results are shown on FIGS. 13(A) and 13(B), where it can be seenthat i.p. treatment with the nucleoside significantly reduced Aβ40 andAβ42 plaque burden as well as the size distribution of remaining plaquesafter seven days of treatment. All studies were performed blind totreatment condition.

In addition to testing in rodent models, as described above, compoundsof the present disclosure may be tested in canine models for humanneurodegenerative diseases, such as dogs with canine counterpart ofAlzheimer's disease. Further, ADME (Absorption, Distribution, Metabolismand Excretion) and Toxicity (ADMET) Studies for safety, tolerability,and pharmacokinetic (PK) profiles of the compounds of the presentdisclosure are conducted in rodents and a second species (such as dogs).For example, an aged beagle model is available from InterVivo (see, thewebsite on the world wide web intervivo.com/aged-dog/ad). Other modelshave been reported in the literature, such as by Insua et al., NeurobiolAging, 2010, 31(4): 625-635 (epub 2008 Jun. 24, doi:10.1016/j.neurobiolaging.2008.05.014).

Example 14 Oral Bioavailability of Compound 5

The mean value of oral bioavailability of the nucleoside compound 5 is72.2%. In human whole blood compound 5 has a half-life of 982 minutes.

Example 15 Administration of the Diphosphate Derivative of Compound 5Decreased Levels of Circulating Cytokines in the Plasma of PSAPP Mice

Inflammatory cytokines were assessed in mouse plasma of both wild typeand PS1/APP mice (Alzheimer's mouse model) and the impact ofintraperitoneal delivery of the diphosphate derivative of compound 5 oncirculating cytokines was assessed. Seven daily intraperitonealinjections of 1 μg/kg of the diphosphate derivative of compound 5 weredelivered to mice. 24 hours following the final injection, plasma wascollected. Wildtype, age matched littermates and PSAPP mice (>6 monthsof age) were treated either with vehicle (phosphate buffered saline) orvehicle containing compound. Compared to wildtype mice, PSAPP miceexhibited greater levels of several cytokines (where cytokine levelswere measured in pg/ml. In particular IL-9, IL-15 and MIG were elevatedcompared to WT vehicle controls. Treatment with compound reduced thelevels of several cytokines in PSAPP mice including Il-1β, IL-2, IL-7,IL-9, IL-10, IL-15, MIG and MIP1α. Additionally, treatment reduced thelevels of certain cytokines in wildtype animals (e.g. IL-2 and IL-10)supporting the conclusion that agonizing P₂Y₆ receptor activity,directly or indirectly, affects inflammatory cytokines more generally(e.g., not specifically in Alzheimers models). See FIG. 14.

Examples 16 Administration of Compound 5 to Pre-Symptomatic PSAPP MiceReduced Amyloid β Accumulation and Reduced Impairment

In this study, treatment of PSAPP mice commenced when the mice werepre-symptomatic. Specifically, treatment was initiated when the micewere approximately 3 months old. PSAPP mice were treated for 100 dayswith daily, intraperitoneal injections of 10 μg/kg of compound 5 orvehicle. Following treatment, mice were assessed in a fear conditioningtask for memory formation. Subsequently, plaque burden in the mice wasalso evaluated. The data provided in Examples 16 and 17 is for 19vehicle treated mice and 22 compound treated mice. The data provided forthe vehicle-treated group is the average across those mice, and errorbars depict the standard error (SEM). Similarly, the data provided forthe compound-treated group is the average across those mice, and theerror bars depict the SEM.

Treatment with compound 5 prevented impairments in contextual fearmemory. In other words, treatment decreased loss of memory thatotherwise develops and is observed in PSAPP mice. Memory is assessed bythe percent time that mice “freeze” 24 hours after a fear inducingelectric shock. The greater the freezing the greater the memory of theprior shock. As depicted in FIG. 15, the compound 5 treated mice show astatistically significant increase in this freezing time, as compared tovehicle treated mice.

In addition, mice treated with compound 5 had reduced plaque burden.This reflects a reduction in amyloid β accumulation in the compound 5treated mice versus the vehicle treated mice. See FIG. 16. In FIGS. 15and 16, compound 5 is indicated along the x-axis as “compound”.

Example 17 Administration of Compound 5 Decreased Levels of Cytokines inthe Plasma of PSAPP Mice

As part of the study described in Example 16, the levels of numerouscytokines in plasma of mice treated with vehicle or compound 5 were alsoevaluated. As described in Example 16, treatment was initiated when themice were approximately 3 months old. PSAPP mice were treated for 100days with daily injections of 10 μg/kg of compound 5 or vehicle.Following treatment and the fear conditioning task experiment, plasmawas taken for multiplex cytokine analysis of circulating cytokine levelsin vehicle-treated versus compound-treated PSAPP mice. Cytokine levelsare assayed using a multi-plex system where beads are labelled withcapture antibody specific for each analyte tested. Each bead set iscoupled to a specific capture antibody and is distinguishable from beadscoupled to a different capture antibody. Thus, the levels of eachanalyte can be evaluated and distinguished. These results are summarizedin FIGS. 17-20, and levels of each analyte are shown in pg/ml. In FIGS.17-20, compound 5 is indicated along the x-axis as “compound”.

Briefly, following treatment for 100 days with compound 5, astatistically significant reduction, in plasma levels of IL-4, IL-10,and IL-12 (also referred to as IL-12(p70)) was observed, in comparisonto vehicle control treated mice. See FIG. 17. For IL-12, the reductionobserved represents an actual reduction in the functional IL-12,heterodimeric cytokine, and not just a reduction in the p40 subunitcommon to multiple cytokines. Specifically, the data examining the p40subunit alone (depicted as IL-12(p40); using a capture antibody thatmeasures the p40 subunit) does not reflect a change following treatmentwith compound 5 while the IL-12(p70) data shows a statisticallysignificant reduction following treatment with compound 5 (see FIG. 17).Regardless of the mechanism of action by which IL-12 is reduced, theseresults indicate that it is not merely via a mechanism generic to allcytokines that share the p40 subunit. Throughout the application, werefer to a reduction in IL-12 cytokine levels interchangeably byreference to either “IL-12” or “IL-12(p70)”.

In addition, the average plasma levels of numerous other cytokines, suchas IFN-r, IL-1β, IL-2, IL-3, IL-5, IL-6, IL-7, IL-9, IL-13, IL-17, LIF,MIP-1α, and MIP-1β, were also reduced. However, administration of thecompound did not result in overall immunosuppression, as the levels ofseveral cytokines were not changed in treated mice. For example, no orsubstantially no change in the levels of M-CSF, MIP-2, Eotaxin, GM-CSF,G-CSF, LIX, MCP-1, IL-1α and IP-10 was observed.

Example 18 Administration of Compound 5 Reduced Amyloid β, PreventedMemory Deficit and Reduced Levels of Pro-Inflammatory Cytokines in PSAPPMice

In this study, treatment of PSAPP mice commenced when the mice werepre-symptomatic. Specifically, treatment was initiated when the micewere approximately 3 months old. PSAPP mice were treated for 100 dayswith daily, intraperitoneal injections of 10 μg/kg of compound 5 orvehicle. Following treatment, mice were assessed for the following:concentration of circulating pro-inflammatory cytokines; amyloid betaload (e.g., plaque burden); and memory in a fear conditioning task formemory formation. The data provided in Example 18 is for 18 vehicletreated mice and 20 compound treated mice. The data provided for thevehicle-treated group is the average across those mice, and error barsdepict the standard error (SEM). Similarly, the data provided for thecompound-treated group is the average across those mice, and the errorbars depict the SEM. Mice were first evaluated in the fear conditioningtask experiment. Subsequently, plasma was taken for multiplex cytokineanalysis of circulating levels of cytokine in the plasma. Finally,plaque burden was assessed in the cortex and hippocampus of the mice.

As depicted in FIG. 21, following treatment for 100 days with compound5, a statistically significant reduction in plasma levels of thefollowing cytokines was observed: IL-12 (also referred to asIL-12(p70)), IL-13, IL-17, IL-10, IL-4, MIP-1a, MIP-1b, and IL-2. SeeFIG. 21. For IL-12, the reduction observed represents an actualreduction in the functional IL-12, heterodimeric cytokine, and not justa reduction in the p40 subunit common to multiple cytokines, asexplained above. For each cytokine examined, plasma levels (e.g., plasmaconcentration) are measured in pg/ml and the observed levels in compound5 treated mice are compared to those observed in vehicle control treatedmice.

As depicted in FIG. 22(C), treatment with compound 5 preventedimpairments in contextual fear memory. In other words, treatmentdecreased loss of memory that otherwise develops and is observed inPSAPP mice. Memory is assessed by the percent time that mice “freeze” 24hours after a fear inducing electric shock. The greater the freezing thegreater the memory of the prior shock. As depicted in FIG. 22(C), thecompound 5 treated mice show a statistically significant increase inthis freezing time, as compared to vehicle treated mice.

In addition, mice treated with compound 5 had reduced plaque burden.This reflects a reduction in amyloid β accumulation in the compound 5treated mice versus the vehicle treated mice (See FIGS. 22(A) and22(B)). Amyloid beta accumulation was decreased in both the cortex andthe hippocampus of compound 5 treated mice. See FIGS. 22(A) and 22(B),respectively. In FIG. 21, the asterisks correspond to the following pvalues: * p<0.05; ** p<0.01. In FIG. 22, the asterisks correspond to thefollowing p values: for 22A, *** p<0.01; for 22B and C, * p<0.02.

Example 19 Seven Day Treatment with Compound 5 Reversed Amyloid βAccumulation and Reduced Levels of Pro-Inflammatory Cytokines in PSAPPMice

In this study, treatment of PSAPP mice commenced after the mice werealready symptomatic. Specifically, treatment was initiated when the micewere approximately 6 months old. PSAPP mice were treated for 7 days withdaily, intraperitoneal injections of 10 μg/kg of compound 5 or vehicle.Following treatment, mice were assessed for the following: concentrationof circulating pro-inflammatory cytokines; amyloid beta load (e.g.,plaque burden); and plaque size. The data provided in Example 19 is for7 vehicle treated mice and 7 compound treated mice. The data providedfor the vehicle-treated group is the average across those mice, anderror bars depict the standard error (SEM). Similarly, the data providedfor the compound-treated group is the average across those mice, and theerror bars depict the SEM. Plasma was taken for multiplex cytokineanalysis of circulating levels of cytokine in the plasma. Subsequently,plaque burden was assessed in the cortex (using Aβ40) and hippocampus(using Aβ42) of the mice, and plaque size was also assessed.

As depicted in FIG. 23, following treatment for 7 days with compound 5,a statistically significant reduction in plasma levels of the followingcytokines was observed: IL-12 (also referred to as IL-12(p70)), IL-13,IL-17, IL-10, MIP-1a, MIP-1b, and TNFα. See FIG. 23. For each cytokineexamined, plasma levels (e.g., plasma concentration) were measured inpg/ml and the observed levels in compound 5 treated mice were comparedto those observed in vehicle control treated mice.

As depicted in FIGS. 24(A) and 24(B), mice treated with compound 5 hadreduced plaque burden. This reflects a reduction in amyloid βaccumulation in the compound 5 treated mice versus the vehicle treatedmice (See FIG. 24(A)). Amyloid beta accumulation was decreased in boththe cortex (upper panel of FIG. 24(A)) and the hippocampus (lower panelof FIG. 24(A)) of compound 5 treated mice. In addition, a decrease inplaque size was was observed in compound 5 treated mice (See FIG.24(B)). In FIG. 23, the asterisks correspond to the following pvalues: * p<0.05; ** p<0.005; *** p<0.0005. In FIG. 24, the asteriskscorrespond to the following p values: * p<0.05.

Example 20 P₂Y₆ Modulating Compounds Mediate Cytokine Release in HumanTHP-1 Cells

THP-1 cells, a human cell line derived from monocytes, were treated invitro with vehicle, the P₂Y₆ receptor antagonist MRS 2578, or compound5, singly or in combination. THP-1 cells were seeded in 24 well platesat a density of 4×10⁵ cells/well in culture medium for 24 hours.Thereafter, vehicle, compound 5, or MRS 2578 (singly or in combination)were added to the cultures for 24 hours. Subsequently the culture mediumwas aspirated and subjected to multiplex cytokine analysis. Data arepresented as means+/−sem. See FIGS. 25 and 26. MRS 2578 has a molecularweight of about 472 and is also described using CAS number 711019-86-2.

As summarized in FIG. 25, treatment of human THP-1 cells with the P₂Y₆receptor antagonist MRS 2578 increased the release of certain cytokinesfrom the cells, as measured by an increase in the observed concentrationof particular cytokines in the culture medium, consistent with ananti-inflammatory effect endogenously exerted by this receptor. Numerouscytokines were evaluated, and data for fractalkine, PDGF-BB, and IL-7are shown in FIG. 25. Observed concentration of these cytokines in theculture medium, following treatment, is indicated along the y-axis inpg/ml. In FIG. 25, the asterisks correspond to the following p values: *p<0.05; ** p<0.02; *** p<0.002.

Moreover, our studies in human THP-1 cells showed that the effects ofmodulating P₂Y₆ receptor activity were reciprocal. Specifically,treating cells in culture with compound 5 (10 nM) reduced the release ofcytokines. This is consistent with the inhibitory affects on plasmacytokine concentration observed following administration of compound 5to mice. This inhibitory affect was blocked by co-incubating the cellsin both compound 5 and the P₂Y₆ receptor antagonist MRS 2578. FIG. 26summarizes the results of an experiment in which IL-7 release from humanTHP-1 cells was examined.

Interestingly, the concentration of compound 5 used in this experiment,10 nM, is in good agreement with the effective dose of compound 5 (10ug/kg) that gives a Cmax of 13 nM.

FIG. 26 summarizes data evaluating concentration of cytokine (IL-7)released from human THP-1 cells treated in vitro with vehicle, compound5, the P₂Y₆ antagonist MRS 2578, or both compound 5 and MRS 2578.Concentration of cytokine secreted into the culture media is measured inpg/ml, as represented on the y-axis. Treatment with MRS 2578 increasedrelease of IL-7 from human THP-1 cells. Treatment with compound 5decreased release of IL-7 from human THP-1 cells, and this effect wasabrogated by co-treatment with MRS 2578. In FIG. 26, the asteriskscorrespond to the following p values: * p<0.05.

What is claimed is:
 1. A pharmaceutical composition, comprising acompound of formula I:

or a salt thereof, wherein: A is a bicyclic aromatic ring selected from:

wherein the bicyclic aromatic ring is independently and optionallysubstituted with one or more R⁷; X is selected from: —H, —C(O)R⁵, and—C(O)OR⁵; Y is a bond or a (C1-C5)-aliphatic group independently andoptionally substituted with one or more R⁴; Z and W are eachindependently selected from ═O, ═S, ═N(R⁵), and ═NOR⁵; R¹ is selectedfrom: —H, bromine, iodine, methyl, ethyl or —CF₃; R² and R³ are eachindependently selected from —OR⁵, —SR⁵, —NR⁵R⁶, —OC(O)R⁵, —OC(O)NR⁵R⁶,and —OC(O)OR⁵; each occurrence of R⁴ is independently selected from:halogen, —OR⁵, —NO₂, —CN, —CF₃, —OCF₃, —R⁵, 1,2-methylenedioxy,1,2-ethylenedioxy, —N(R⁵)₂, —SR⁵, —SOR⁵, —SO₂R⁵, —SO₂N(R⁵)₂, —SO₃R⁵,—C(O)R⁵, —C(O)C(O)R⁵, —C(O)CH₂C(O)R⁵, —C(S)R⁵, —C(S)OR⁵, —C(O)OR⁵,—C(O)C(O)OR⁵, —C(O)C(O)N(R⁵)₂, —OC(O)R⁵, —C(O)N(R⁵)₂, —OC(O)N(R⁵)₂,—C(S)N(R⁵)₂, —(CH₂)₀₋₂NHC(O)R⁵, —N(R⁵)N(R⁵)COR⁵, —N(R⁵)N(R⁵)C(O)OR⁵,—N(R⁵)N(R⁵)CON(R⁵)₂, —N(R⁵)SO₂R⁵, —N(R⁵)SO₂N(R⁵)₂, —N(R⁵)C(O)OR⁵,—N(R⁵)C(O)R⁵, —N(R⁵)C(S)R⁵, —N(R⁵)C(O)N(R⁵)₂, —N(R⁵)C(S)N(R⁵)₂,—N(COR⁵)COR⁵, —N(OR⁵)R⁵, —C(═NH)N(R⁵)₂, —C(O)N(OR⁵)R⁵, —C(═NOR⁵)R⁵,—OP(O)(OR⁵)₂, —P(O)(R⁵)₂, —P(O)(OR⁵)₂, and —P(O)(H)(OR⁵); eachoccurrence of R⁵ is independently selected from: H—,(C1-C12)-aliphatic-, (C3-C10)-cycloalkyl- or -cycloalkenyl-,[(C3-C10)-cycloalkyl or -cycloalkenyl]-(C1-C12)-aliphatic-,(C6-C10)-aryl-, (C6-C10)-aryl-(C1-C12)aliphatic-,(C3-C10)-heterocyclyl-, (C6-C10)-heterocyclyl-(C1-C12)aliphatic-,(C5-C10)-heteroaryl-, and (C5-C10)-heteroaryl-(C1-C12)-aliphatic-;wherein two R⁵ groups bound to the same atom optionally form a 3- to10-membered aromatic or non-aromatic ring having up to 3 heteroatomsindependently selected from N, O, S, SO, and SO₂, wherein said ring isoptionally fused to a (C6-C10)aryl, (C5-C10)heteroaryl,(C3-C10)cycloalkyl, or a (C3-C10)heterocyclyl; and wherein each R⁵ groupis independently and optionally substituted with one or more R⁷; R⁶ isselected from: —R⁵, —C(O)R⁵, —C(O)OR⁵, —C(O)N(R⁵)₂ and —S(O)₂R⁵; eachoccurrence of R⁷ is independently selected from: halogen, —OR⁸, —NO₂,—CN, —CF₃, —OCF₃, —R⁸, oxo, thioxo, 1,2-methylenedioxy,1,2-ethylenedioxy, —N(R⁸)₂, —SR⁸, —SOR⁸, —SO₂R⁸, —SO₂N(R⁸)₂, —SO₃R⁸,—C(O)R⁸, —C(O)C(O)R⁸, —C(O)CH₂C(O)R⁸, —C(S)R⁸, —C(S)OR⁸, —C(O)OR⁸,—C(O)C(O)OR⁸, —C(O)C(O)N(R⁸)₂, —OC(O)R⁸, —C(O)N(R⁸)₂, —OC(O)N(R⁸)₂,—C(S)N(R⁸)₂, —(CH₂)₀₋₂NHC(O)R⁸, —N(R⁸)N(R⁸)COR⁸, —N(R⁸)N(R⁸)C(O)OR⁸,—N(R⁸)N(R⁸)CON(R⁸)₂, —N(R⁸)SO₂R⁸, —N(R⁸)SO₂N(R⁸)₂, —N(R⁸)C(O)OR⁸,—N(R⁸)C(O)R⁸, —N(R⁸)C(S)R⁸, —N(R⁸)C(O)N(R⁸)₂, —N(R⁸)C(S)N(R⁸)₂,—N(COR⁸)COR⁸, —N(OR⁸)R⁸, —C(═NH)N(R⁸)₂, —C(O)N(OR⁸)R⁸, —C(═NOR⁸)R⁸,—OP(O)(OR⁸)₂, —P(O)(R⁸)₂, —P(O)(OR⁸)₂, and —P(O)(H)(OR⁸); eachoccurrence of R⁸ is independently selected from: H— and(C1-C6)-aliphatic-; and an acceptable carrier, adjuvant or excipient. 2.The pharmaceutical composition of claim 1, wherein X of the compound offormula I is —H or —C(O)R⁵.
 3. The pharmaceutical composition of claim1, wherein R¹ is —H.
 4. The pharmaceutical composition of claim 1,wherein Z of the compound of formula I is ═O or ═S.
 5. Thepharmaceutical composition of claim 1, wherein Z is ═O.
 6. Thepharmaceutical composition of claim 1, wherein W of the compound offormula I is ═O or ═S.
 7. The pharmaceutical composition of claim 1,wherein W is ═O.
 8. The pharmaceutical composition of claim 1, wherein Yof the compound of formula I is a C1-aliphatic group optionallysubstituted with one or more R⁴.
 9. The pharmaceutical composition ofclaim 1, wherein Y is —CH₂—.
 10. The pharmaceutical composition of claim1, wherein Y of the compound of formula I is a C2-aliphatic groupoptionally substituted with one or more R⁴.
 11. The pharmaceuticalcomposition of claim 1, wherein Y is —CH₂—C(R⁴)₂—.
 12. Thepharmaceutical composition of claim 1, wherein Y is —CH₂—CH₂—.
 13. Thepharmaceutical composition of claim 1, wherein each occurrence of R⁴ isindependently selected from halogen.
 14. The pharmaceutical compositionof claim 1, wherein both occurrences of R⁴ are —F.
 15. Thepharmaceutical composition of claim 1, wherein each occurrence of R⁴ isindependently a (C1-C3)-aliphatic group.
 16. The pharmaceuticalcomposition of claim 1, wherein both occurrences of R⁴ are —CH₃.
 17. Thepharmaceutical composition of claim 1, wherein R² of the compound offormula I is —OR⁵.
 18. The pharmaceutical composition of claim 1,wherein R² is —OH.
 19. The pharmaceutical composition of claim 1,wherein R³ of the compound of formula I is —OR⁵.
 20. The pharmaceuticalcomposition of claim 1, wherein R³ is —OH.